LOCUS       sequence-112594-       13743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  const. Mis12-GFP-FKBP3; ind. FRB-mCH-Mad1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13743)
  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 13743)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2014-03-17- 17"; volume: "204"; 
            issue: "6"; pages: "901-8"
COMMENT     Sequence Label: pERB130
FEATURES             Location/Qualifiers
     source          1..13743
                     /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           591..632
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     regulatory      1392..1401
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1398..1400,1401..1403,1404..2114)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1398..1400,1401..1403,1404..2114)
                     /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(1443..1464)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1704..1723)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2051..2072
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2133..2453
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2460..2780
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /note="synthetic gene with alternative codons"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2460..2780
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2793..3116
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(3223..3797,3798..3800,3801..3809)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3390..3407)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3617..3636
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3810..4409
                     /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(3810..3829)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4306..4326
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4524..4541)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4530..4641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4737..4858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4990..5697)
                     /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(5708..6919)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5852..5871)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    6941..6959
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6976..6994
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7012..7030
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7048..7066
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7083..7101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7119..7137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        7149..7187
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7184..7208
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     ncRNA           7365..7391
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           7417..7458
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     CDS             7599..7877
                     /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"
     regulatory      7890..7899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7896..8603
                     /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(8041..8059)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8309..8328
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(10890..10909)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    10955..11010
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(11009..11028)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    11371..11404
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    11417..11538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11557..11577)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11557..11575)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11596..11612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11596..11612)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11609..11631)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11620..11636
                     /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(11644..11650,11651..11668,11669..11674))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11689..11710
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11827..11844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11998..12586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12078..12097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12757..13548,13549..13617))
                     /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     13380..13399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13618..13722)
                     /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 gtatgctgat atggattcat
      541 gtgcacatgg ttttggccac tgactgacca tgtgcatgaa tccatatcag gacacaaggc
      601 ctgttactag cactcacatg gaacaaatgg ccgaattcgg cgctagctgc tgatatcgca
      661 tacgcgtgga ccagataggc acctattggt cttactgaca tccactttgc ctttctctcc
      721 acaggtgtcg gtaccgcggg cccgggatcc accggcgcgc caccatgtct gtggatccaa
      781 tgacctacga ggcccagttc tttggcttca cgccacaaac gtgcatgctt cggatctaca
      841 ttgcatttca agactaccta tttgaagtga tgcaggccgt tgaacaggtt attctgaaga
      901 agctggatgg catcccagac tgtgacatta gcccagtgca gattcgcaaa tgcacagaga
      961 agtttctttg cttcatgaaa ggacattttg ataacctttt tagcaaaatg gagcaactgt
     1021 ttttgcagct gattttacgt attccctcaa acatcttgct tcctgaagat aaatgtaagg
     1081 agacacctta tagtgaggaa gattttcagc atctccagaa agaaattgaa cagttacagg
     1141 agaagtacaa gactgaatta tgtactaagc aggcccttct tgcagaatta gaagagcaaa
     1201 aaattgttca ggccaaactc aaacagacgt tgactttctt tgatgagctt cataatgttg
     1261 gcagagatca tgggactagt gattttaggg agagtttagt atccctggtt cagaactcca
     1321 gaaaactaca gaacattaga gacaatgtgg aaaaggaatc gaaacgactg aaaatatcta
     1381 tgctagcagt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1441 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1501 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1561 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     1621 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1681 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1741 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1801 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1861 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     1921 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1981 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     2041 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2101 acgagctgta caagggtggt gaattctcta gaggagtgca ggtggaaacc atctccccag
     2161 gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc
     2221 ttgaagatgg aaagaaattt gattcctccc gggacagaaa caagcccttt aagtttatgc
     2281 taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc
     2341 agagagccaa actgactata tctccagatt atgcctatgg tgccactggg cacccaggca
     2401 tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg gaaactagag
     2461 gcgtgcaggt cgagaccatc agccccggcg acggccgcac ctttcccaag agaggccaga
     2521 cttgcgtggt ccactacacc ggcatgctgg aggacggcaa gaagttcgac agcagccgcg
     2581 accgcaacaa gcccttcaag ttcatgctgg gcaaacagga agtgatccgc ggctgggagg
     2641 aaggcgtggc tcagatgagc gtggggcagc gggccaagct gaccatcagc cccgactatg
     2701 cctacggcgc caccggccac cccggcatca tcccccccca cgccaccctg gtgttcgacg
     2761 tggagctgct gaagctggag actagtgaat tcggagtgca ggtggaaacc atctccccag
     2821 gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc
     2881 ttgaagatgg aaagaaattt gattcctccc gggacagaaa caagcccttt aagtttatgc
     2941 taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc
     3001 agagagccaa actgactata tctccagatt atgcctatgg tgccactggg cacccaggca
     3061 tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg gaatgatcta
     3121 gagggcccta ttctccggat aaccggaggt gcaggctgcc tatgcatgtt aacaattgca
     3181 tcgataagct tgatatcgaa ttcctgcagc ccgggggatc cgcccctctc cctccccccc
     3241 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     3301 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     3361 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     3421 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     3481 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     3541 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     3601 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     3661 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt
     3721 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     3781 aaaaacacga tgataatatg gccacaacca tgaccgagta caagcccacg gtgcgcctcg
     3841 ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc
     3901 ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag
     3961 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4021 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4081 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     4141 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtgt
     4201 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     4261 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     4321 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     4381 ggtgcatgac ccgcaagccc ggtgcctagc cggagaccga aaggagcgca cgaccccatg
     4441 gctccgaccg aagccgaccc gggcggcccc gccgaccccg cacccgcccc cgaggcccac
     4501 cgactctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4561 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4621 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4681 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcactagtt tcgcgttaag
     4741 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4801 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4861 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     4921 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagttat ctagatccgg
     4981 tggatctatt tacccgggga gcatgtcaag gtcaaaatcg tcaagggcat cggctgggag
     5041 catgtctaag tcaaaatcgt caagggcgtc ggtcggcccg ccgctttcgc actttagctg
     5101 tttctccagg ccacatatga ttagttccag gccgaaaagg aaggcaggtt cggctccctg
     5161 ccggtcgaac agctcaattg cttgtctcag aagtgggggc atagaatcgg tggtaggtgt
     5221 ctctctttcc tcttttgcta cttgatgctc ctgttcctcc aatacgcagc ccagtgtaaa
     5281 gtggcccacg gcggacagag cgtacagtgc gttctccagg gagaagcctt gctgacacag
     5341 gaacgcgagc tgattttcca gggtttcgta ctgtttctct gttgggcggg tgccgagatg
     5401 cactttagcc ccgtcgcgat gtgagaggag agcacagcgg tatgacttgg cgttgttccg
     5461 cagaaagtct tgccatgact cgccttccag ggggcagaag tgggtatgat gcctgtccag
     5521 catctcgatt ggcagggcat cgagcagggc ccgcttgttc ttcacgtgcc agtacagggt
     5581 aggctgctca actcccagct tttgagcgag tttccttgtc gtcaggcctt cgataccgac
     5641 accattgagt aattccagag ctccgtttat gactttgctc ttgtccagcc tagacatggt
     5701 ggctagcgtc taacaaaaaa gccaaaaacg gccagaattt agcggacaat ttactagtct
     5761 aacactgaaa attacatatt gacccaaatg attacatttc aaaaggtgcc taaaaaactt
     5821 cacaaaacac actcgccaac cccgagcgca tagttcaaaa ccggagcttc agctacttaa
     5881 gaagataggt acataaaacc gaccaaagaa actgacgcct cacttatccc tcccctcacc
     5941 agaggtccgg cgcctgtcga ttcaggagag cctaccctag gcccgaaccc tgcgtcctgc
     6001 gacggagaaa agcctaccgc acacctaccg gcaggtggcc ccaccctgca ttataagcca
     6061 acagaacggg tgacgtcacg acacgacgag ggcgcgcgct cccaaaggta cgggtgcact
     6121 gcccaacggc accgccataa ctgccgcccc cgcaacagac gacaaaccga gttctccagt
     6181 cagtgacaaa cttcacgtca gggtccccag atggtgcccc agcccatctc acccgaataa
     6241 gagctttccc gcattagcga aggcctcaag accttgggtt cttgccgccc accatgcccc
     6301 ccaccttgtt tcaacgacct cacagcccgc ctcacaagcg tcttccattc aagactcggg
     6361 aacagccgcc attttgctgc gctcccccca acccccagtt cagggcaacc ttgctcgcgg
     6421 acccagacta cagcccttgg cggtctctcc acacgcttcc gtcccaccga gcggcccggc
     6481 ggccacgaaa gccccggcca gcccagcagc ccgctactca ccaagtgacg atcacagcga
     6541 tccacaaaca agaaccgcga cccaaatccc ggctgcgacg gaactagctg tgccacaccc
     6601 ggcgcgtcct tatataatca tcggcgttca ccgccccacg gagatccctc cgcagaatcg
     6661 ccgagaaggg actacttttc ctcgcctgtt ccgctctctg gaaagaaaac cagtgcccta
     6721 gagtcaccca agtcccgtcc taaaatgtcc ttctgctgat actggggttc taaggccgag
     6781 tcttatgagc agcgggccgc tgtcctgagc gtccgggcgg aaggatcagg acgctcgctg
     6841 cgcccttcgt ctgacgtggc agcgctcgcc gtgaggaggg gggcgcccgc gggaggcgcc
     6901 aaaacccggc gcggaggcca cgcgtactag acgagtttac tccctatcag tgatagagaa
     6961 cgatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta ctccctatca
     7021 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgagtt
     7081 tatccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
     7141 tatgtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc
     7201 gtcagatcgc accgggtgac tctcttaagg tagccttgca gaagttggtc gtgaggcact
     7261 gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     7321 tcgagacaga gaagactctt gcgtttcggt cgacttctag agctctggag gcttgctgaa
     7381 ggctgtatgc tagagacgta cagatgcgtc tcacaggaca caaggcctgt tactagcact
     7441 cacatggaac aaatggccga attcggcgct agctgctgat atcgcatacg cgtggaccag
     7501 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtcggtac
     7561 cgcgggcccg ggatccaccg gtcaccatgg cttctagaat cctctggcat gagatgtggc
     7621 atgaaggcct ggaagaggca tctcgtttgt actttgggga aaggaacgtg aaaggcatgt
     7681 ttgaggtgct ggagcccttg catgctatga tggaacgggg cccccagact ctgaaggaaa
     7741 catcctttaa tcaggcctat ggtcgagatt taatggaggc ccaagagtgg tgcaggaagt
     7801 acatgaaatc agggaatgtc aaggacctcc tccaagcctg ggacctctat tatcatgtgt
     7861 tccgacgaat ctcaaagact agtccggtcg ccaccatggt gagcaagggc gaggaggata
     7921 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     7981 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     8041 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     8101 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     8161 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     8221 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     8281 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     8341 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     8401 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     8461 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     8521 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     8581 gcggcatgga cgagctgtat aaggccagcc tgatggaaga cctgggggaa aacaccatgg
     8641 ttttatccac cctgagatct ttgaacaact tcatctctca gcgtgtggag ggaggctctg
     8701 gactggatat ttctacctcg gccccaggtt ctctgcagat gcagtaccag cagagcatgc
     8761 agctggagga aagagcagag cagatccgtt cgaagtccca cctcatccag gtggagcggg
     8821 agaaaatgca gatggagctg agtcacaaga gggctcgagt ggagctggag agagcagcca
     8881 gcaccagtgc caggaactac gagcgtgagg tcgaccgcaa ccaggagctc ctgacgcgca
     8941 tccggcagct tcaggagcgg gaggccgggg cggaggagaa gatgcaggag cagctggagc
     9001 gcaacaggca gtgtcagcag aacttggatg ctgccagcaa gaggctgcgt gagaaagagg
     9061 acagtctggc ccaggctggc gagaccatca acgcactgaa ggggaggatc tcggaactgc
     9121 agtggagcgt gatggaccag gagatgcggg tgaagcgcct ggagtcggag aagcaggagc
     9181 tgcaggagca gctggacctg caacacaaaa aatgccagga agccaatcag aaaatccagg
     9241 aactccaggc cagccaagaa gcaagagcag accacgagca gcagattaag gatctggagc
     9301 agaagctgtc cctgcaagag caggatgcag cgattgtgaa gaacatgaag tctgagctgg
     9361 tacggctccc taggctggaa cgggagctga agcagctgcg ggaggagagc gcgcacctgc
     9421 gggaggagag cgcgcacctg cgggagatga gagagaccaa cgggctgctc caggaagagc
     9481 tggaagggct gcagaggaag ctggggcgcc aggagaagat gcaggagacg ctggttggct
     9541 tggagctgga gaacgagagg ctgctggcca agctgcaaag ctgggagaga ctggaccaga
     9601 ccatgggcct gagcatcagg actccagaag acctttccag attcgtggtt gagctgcagc
     9661 agagggagct tgccttgaag gacaagaaca gcgccgtcac cagcagcgcc cgggggctgg
     9721 agaaggccag gcagcagctg caggaggagc tccggcaggt cagcggccag ctgttggagg
     9781 agaggaagaa gcgcgagacc cacgaggcgc tggcccggag gctccagaaa cgggtcctgc
     9841 tgctcaccaa ggagcgggac ggtatgcggg ccatcctggg gtcctacgac agcgagctga
     9901 ccccggccga gtactcaccc cagctgacgc ggcgcatgcg ggaggctgag gatatggtgc
     9961 agaaggtgca cagccacagc gccgagatgg aggctcagct gtcgcaggcc ctggaggagc
    10021 tgggaggcca gaaacaaaga gcagacatgc tggagatgga gctgaagatg ctgaagtctc
    10081 agtccagctc tgccgaacag agcttcctgt tctccaggga ggaggcggac acgctcaggt
    10141 tgaaggtcga ggagctggaa ggcgagcgga gtcggctgga ggaggaaaag aggatgctgg
    10201 aggcacagct ggagcggcga gctctgcagg gtgactatga ccagagcagg accaaagtgc
    10261 tgcacatgag cctgaacccc accagtgtgg ccaggcagcg cctgcacgag gaccacagcc
    10321 agctgcaggc ggagtgcgag cgactgcgcg ggctcctgcg cgccatggag agaggaggca
    10381 ccgtcccagc cgaccttgag gctgccgccg cgagtctgcc atcgtccaag gaggtggcag
    10441 agctgaagaa gcaggtggag agtgccgagc tgaagaacca gcggctcaag gaggttttcc
    10501 agaccaagat ccaggagttc cgcaaggcct gctacacgct caccggctac cagatcgaca
    10561 tcaccacgga gaaccagtac cggctgacct cgctgtacgc cgagcaccca ggcgactgcc
    10621 tcatcttcaa ggccaccagc ccctcgggtt ccaagatgca gctactggag acagagttct
    10681 cacacaccgt gggcgagctc atcgaggtgc acctgcggcg ccaggacagc atccctgcct
    10741 tcctcagctc gctcaccctc gagctcttca gccgccagac cgtggcgtag ggatccctat
    10801 gatgtacaag taaacactca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
    10861 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
    10921 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
    10981 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
    11041 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccat atgctggctg
    11101 ccatgaacaa aggtggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
    11161 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
    11221 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
    11281 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgaa gatccctcga
    11341 cctgagcgtc gagctagcat atggatccat ataacttcgt ataatgtatg ctatacgaag
    11401 ttattacgat accgtcaact tgtttattgc agcttataat ggttacaaat aaagcaatag
    11461 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
    11521 actcatcaat gtatcttagg tacccagctt ttgttccctt tagtgagggt taattgcgcg
    11581 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
    11641 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
    11701 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
    11761 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
    11821 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
    11881 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
    11941 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
    12001 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
    12061 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
    12121 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
    12181 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
    12241 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
    12301 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
    12361 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
    12421 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
    12481 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
    12541 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
    12601 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
    12661 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
    12721 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
    12781 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
    12841 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
    12901 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
    12961 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
    13021 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
    13081 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
    13141 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
    13201 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
    13261 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
    13321 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
    13381 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
    13441 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
    13501 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
    13561 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
    13621 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    13681 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    13741 gcc
//
