LOCUS       sequence-89741-e       11517 bp    DNA     circular SYN 15-APR-2025
DEFINITION  constitutive miRempty_Mis12-GFP-FKBP3; inducible mCh-FRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11517)
  AUTHORS   Ballister ER, Riegman M, Lampson MA
  TITLE     Recruitment of Mad1 to metaphase kinetochores is sufficient to 
            reactivate the mitotic checkpoint.
  JOURNAL   J Cell Biol. 2014 Mar 17;204(6):901-8. doi: 10.1083/jcb.201311113.
  PUBMED    24637323
REFERENCE   2  (bases 1 to 11517)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2014-03-17- 17"; volume: "204"; 
            issue: "6"; pages: "901-8"
COMMENT     Sequence Label: pERB111
FEATURES             Location/Qualifiers
     source          1..11517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    52..100
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    312..345
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     ncRNA           501..527
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           553..594
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     regulatory      1354..1363
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1360..1362,1363..1365,1366..2076)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1360..1362,1363..1365,1366..2076)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1405..1426)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1666..1685)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2013..2034
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2095..2415
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2422..2742
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /note="synthetic gene with alternative codons"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2422..2742
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2755..3078
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(3185..3759,3760..3762,3763..3771)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3352..3369)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3579..3598
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3772..4371
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3772..3791)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4268..4288
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4486..4503)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4492..4603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4699..4820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4952..5659)
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     promoter        complement(5670..6881)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5814..5833)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    6903..6921
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6938..6956
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6974..6992
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7010..7028
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7045..7063
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7081..7099
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        7111..7149
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7146..7170
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     ncRNA           7327..7353
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           7379..7420
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     CDS             7549..8256
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7694..7712)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7962..7981
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             8290..8568
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     primer_bind     complement(8664..8683)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8729..8784
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8783..8802)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    9145..9178
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    9191..9312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9331..9351)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(9331..9349)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9370..9386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9370..9386)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9383..9405)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9394..9410
                     /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(9418..9424,9425..9442,9443..9448))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9463..9484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9601..9618)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9772..10360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9852..9871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10531..11322,11323..11391))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     11154..11173
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11392..11496)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagcttaata aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
       61 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
      121 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
      181 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
      241 gtcggggcgg caggccctgc catagccact ggccccgtgg gttagggacg gggtccccca
      301 ttcgagaata aataacttcg tataaagtat cctatacgaa gttatcaccc tgcagtacgt
      361 atccggtagc cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca
      421 agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt
      481 ttcggtcgac ttctagagct ctggaggctt gctgaaggct gtatgctaga gacgtacaga
      541 tgcgtctcac aggacacaag gcctgttact agcactcaca tggaacaaat ggccgaattc
      601 ggcgctagct gctgatatcg catacgcgtg gaccagatag gcacctattg gtcttactga
      661 catccacttt gcctttctct ccacaggtgt cggtaccgcg ggcccgggat ccaccggcgc
      721 gccaccatgt ctgtggatcc aatgacctac gaggcccagt tctttggctt cacgccacaa
      781 acgtgcatgc ttcggatcta cattgcattt caagactacc tatttgaagt gatgcaggcc
      841 gttgaacagg ttattctgaa gaagctggat ggcatcccag actgtgacat tagcccagtg
      901 cagattcgca aatgcacaga gaagtttctt tgcttcatga aaggacattt tgataacctt
      961 tttagcaaaa tggagcaact gtttttgcag ctgattttac gtattccctc aaacatcttg
     1021 cttcctgaag ataaatgtaa ggagacacct tatagtgagg aagattttca gcatctccag
     1081 aaagaaattg aacagttaca ggagaagtac aagactgaat tatgtactaa gcaggccctt
     1141 cttgcagaat tagaagagca aaaaattgtt caggccaaac tcaaacagac gttgactttc
     1201 tttgatgagc ttcataatgt tggcagagat catgggacta gtgattttag ggagagttta
     1261 gtatccctgg ttcagaactc cagaaaacta cagaacatta gagacaatgt ggaaaaggaa
     1321 tcgaaacgac tgaaaatatc tatgctagca gtcgccacca tggtgagcaa gggcgaggag
     1381 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1441 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1501 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1561 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1621 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1681 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1741 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1801 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1861 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1921 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     1981 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2041 gccgggatca ctctcggcat ggacgagctg tacaagggtg gtgaattctc tagaggagtg
     2101 caggtggaaa ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc
     2161 gtggtgcact acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga
     2221 aacaagccct ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg
     2281 gttgcccaga tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat
     2341 ggtgccactg ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag
     2401 cttctaaaac tggaaactag aggcgtgcag gtcgagacca tcagccccgg cgacggccgc
     2461 acctttccca agagaggcca gacttgcgtg gtccactaca ccggcatgct ggaggacggc
     2521 aagaagttcg acagcagccg cgaccgcaac aagcccttca agttcatgct gggcaaacag
     2581 gaagtgatcc gcggctggga ggaaggcgtg gctcagatga gcgtggggca gcgggccaag
     2641 ctgaccatca gccccgacta tgcctacggc gccaccggcc accccggcat catccccccc
     2701 cacgccaccc tggtgttcga cgtggagctg ctgaagctgg agactagtga attcggagtg
     2761 caggtggaaa ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc
     2821 gtggtgcact acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga
     2881 aacaagccct ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg
     2941 gttgcccaga tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat
     3001 ggtgccactg ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag
     3061 cttctaaaac tggaatgatc tagagggccc tattctccgg ataaccggag gtgcaggctg
     3121 cctatgcatg ttaacaattg catcgataag cttgatatcg aattcctgca gcccggggga
     3181 tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     3241 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     3301 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     3361 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     3421 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     3481 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     3541 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     3601 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     3661 ggtacacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     3721 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac catgaccgag
     3781 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     3841 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     3901 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     3961 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4021 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4081 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4141 ttcctggcca ccgtcggcgt gtcgcccgac caccagggca agggtctggg cagcgccgtc
     4201 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4261 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4321 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta gccggagacc
     4381 gaaaggagcg cacgacccca tggctccgac cgaagccgac ccgggcggcc ccgccgaccc
     4441 cgcacccgcc cccgaggccc accgactcta gagctcgctg atcagcctcg actgtgcctt
     4501 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4561 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4621 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4681 atagcactag tttcgcgtta agatacattg atgagtttgg acaaaccaca actagaatgc
     4741 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4801 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4861 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
     4921 atgatcagtt atctagatcc ggtggatcta tttacccggg gagcatgtca aggtcaaaat
     4981 cgtcaagggc atcggctggg agcatgtcta agtcaaaatc gtcaagggcg tcggtcggcc
     5041 cgccgctttc gcactttagc tgtttctcca ggccacatat gattagttcc aggccgaaaa
     5101 ggaaggcagg ttcggctccc tgccggtcga acagctcaat tgcttgtctc agaagtgggg
     5161 gcatagaatc ggtggtaggt gtctctcttt cctcttttgc tacttgatgc tcctgttcct
     5221 ccaatacgca gcccagtgta aagtggccca cggcggacag agcgtacagt gcgttctcca
     5281 gggagaagcc ttgctgacac aggaacgcga gctgattttc cagggtttcg tactgtttct
     5341 ctgttgggcg ggtgccgaga tgcactttag ccccgtcgcg atgtgagagg agagcacagc
     5401 ggtatgactt ggcgttgttc cgcagaaagt cttgccatga ctcgccttcc agggggcaga
     5461 agtgggtatg atgcctgtcc agcatctcga ttggcagggc atcgagcagg gcccgcttgt
     5521 tcttcacgtg ccagtacagg gtaggctgct caactcccag cttttgagcg agtttccttg
     5581 tcgtcaggcc ttcgataccg acaccattga gtaattccag agctccgttt atgactttgc
     5641 tcttgtccag cctagacatg gtggctagcg tctaacaaaa aagccaaaaa cggccagaat
     5701 ttagcggaca atttactagt ctaacactga aaattacata ttgacccaaa tgattacatt
     5761 tcaaaaggtg cctaaaaaac ttcacaaaac acactcgcca accccgagcg catagttcaa
     5821 aaccggagct tcagctactt aagaagatag gtacataaaa ccgaccaaag aaactgacgc
     5881 ctcacttatc cctcccctca ccagaggtcc ggcgcctgtc gattcaggag agcctaccct
     5941 aggcccgaac cctgcgtcct gcgacggaga aaagcctacc gcacacctac cggcaggtgg
     6001 ccccaccctg cattataagc caacagaacg ggtgacgtca cgacacgacg agggcgcgcg
     6061 ctcccaaagg tacgggtgca ctgcccaacg gcaccgccat aactgccgcc cccgcaacag
     6121 acgacaaacc gagttctcca gtcagtgaca aacttcacgt cagggtcccc agatggtgcc
     6181 ccagcccatc tcacccgaat aagagctttc ccgcattagc gaaggcctca agaccttggg
     6241 ttcttgccgc ccaccatgcc ccccaccttg tttcaacgac ctcacagccc gcctcacaag
     6301 cgtcttccat tcaagactcg ggaacagccg ccattttgct gcgctccccc caacccccag
     6361 ttcagggcaa ccttgctcgc ggacccagac tacagccctt ggcggtctct ccacacgctt
     6421 ccgtcccacc gagcggcccg gcggccacga aagccccggc cagcccagca gcccgctact
     6481 caccaagtga cgatcacagc gatccacaaa caagaaccgc gacccaaatc ccggctgcga
     6541 cggaactagc tgtgccacac ccggcgcgtc cttatataat catcggcgtt caccgcccca
     6601 cggagatccc tccgcagaat cgccgagaag ggactacttt tcctcgcctg ttccgctctc
     6661 tggaaagaaa accagtgccc tagagtcacc caagtcccgt cctaaaatgt ccttctgctg
     6721 atactggggt tctaaggccg agtcttatga gcagcgggcc gctgtcctga gcgtccgggc
     6781 ggaaggatca ggacgctcgc tgcgcccttc gtctgacgtg gcagcgctcg ccgtgaggag
     6841 gggggcgccc gcgggaggcg ccaaaacccg gcgcggaggc cacgcgtact agacgagttt
     6901 actccctatc agtgatagag aacgatgtcg agtttactcc ctatcagtga tagagaacgt
     6961 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
     7021 gatagagaac gtatgtcgag tttatcccta tcagtgatag agaacgtatg tcgagtttac
     7081 tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg tacggtggga ggcctatata
     7141 agcagagctc gtttagtgaa ccgtcagatc gcaccgggtg actctcttaa ggtagccttg
     7201 cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg
     7261 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttcg gtcgacttct
     7321 agagctctgg aggcttgctg aaggctgtat gctagagacg tacagatgcg tctcacagga
     7381 cacaaggcct gttactagca ctcacatgga acaaatggcc gaattcggcg ctagctgctg
     7441 atatcgcata cgcgtggacc agataggcac ctattggtct tactgacatc cactttgcct
     7501 ttctctccac aggtgtcggt accgcgggcc cgggatccac cggtcaccat ggtgagcaag
     7561 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     7621 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     7681 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     7741 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     7801 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     7861 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     7921 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     7981 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     8041 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     8101 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     8161 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     8221 cgccactcca ccggcggcat ggacgagctg tacaagggta ccgcgggccc gggatccacc
     8281 ggatctagaa tcctctggca tgagatgtgg catgaaggcc tggaagaggc atctcgtttg
     8341 tactttgggg aaaggaacgt gaaaggcatg tttgaggtgc tggagccctt gcatgctatg
     8401 atggaacggg gcccccagac tctgaaggaa acatccttta atcaggccta tggtcgagat
     8461 ttaatggagg cccaagagtg gtgcaggaag tacatgaaat cagggaatgt caaggacctc
     8521 ctccaagcct gggacctcta ttatcatgtg ttccgacgaa tctcaaagac tagataagta
     8581 caagtaaaca ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     8641 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     8701 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     8761 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     8821 catcagaatg agtatttggt ttagagtttg gcaacatatg ccatatgctg gctgccatga
     8881 acaaaggtgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     8941 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     9001 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     9061 ctcctgacta ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacctgag
     9121 cgtcgagcta gcatatggat ccatataact tcgtataatg tatgctatac gaagttatta
     9181 cgataccgtc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     9241 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     9301 caatgtatct taggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     9361 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     9421 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     9481 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     9541 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     9601 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     9661 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     9721 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     9781 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     9841 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     9901 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     9961 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
    10021 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
    10081 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
    10141 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
    10201 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
    10261 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
    10321 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
    10381 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
    10441 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
    10501 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
    10561 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
    10621 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
    10681 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
    10741 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
    10801 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
    10861 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
    10921 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
    10981 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
    11041 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
    11101 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
    11161 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
    11221 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    11281 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    11341 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    11401 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    11461 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgcc
//
