LOCUS       Exported               14585 bp ds-DNA     circular SYN 04-OCT-2018
DEFINITION  Acceptor only control for desmoplakin II tension sensor (F40-based).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14585)
  AUTHORS   Anna-Lena Cost
  TITLE     Direct Submission
  JOURNAL   Exported Oct 4, 2018 from SnapGene 4.2.1
            http://www.snapgene.com
COMMENT     pLPCXmod + human desmoplakin II-[YPet(short)(Y67G)-F40-mCherry] 
            (internal-1353)
            
            acceptor only control to co-expressed with donor only control and 
            determine intermolecular FRET
FEATURES             Location/Qualifiers
     source          1..14585
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..589
                     /label=5' LTR
                     /note="5' MoMuSV LTR (long terminal repeat from Moloney 
                     murine sarcoma virus)"
                     /note="color: #f0e1e1"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
                     /note="color: #ffe4c4"
     misc_feature    659..1468
                     /label=-Psi(+)
                     /note="extended packaging signal"
                     /note="color: #f0e1e1"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus gag gene; 
                     lacking the start codon"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    1504..1509
                     /label=AgeI
                     /note="AgeI recognition site"
                     /note="color: #0080ff"
     CDS             1565..2164
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /note="color: #ccffcc"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    1673..1678
                     /label=SalI
                     /note="SalI recognition site"
                     /note="color: #0080ff"
     enhancer        2339..2642
                     /label=CMV enhancer
                     /note="human cytomegalovirus (CMV) immediate early 
                     enhancer"
                     /note="color: #ffe4c4"
     promoter        2643..2846
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffe4c4; direction: RIGHT"
     primer_bind     2843..2867
                     /label=pLPCX_seq_f
                     /note="pLPCX_seq_f"
                     /note="color: blue; sequence: AGCTcGTTTAGTGAACCGTCAGATC"
     misc_feature    2887..2892
                     /label=BglII
                     /note="BglII recognition site"
                     /note="color: #0080ff"
     misc_feature    2912..2917
                     /label=ECoRI
                     /note="ECoRI recognition site"
                     /note="color: #0080ff"
     CDS             join(2921..6979,8519..11275)
                     /codon_start=1
                     /label=Desmoplakin II
                     /note="Human Desmoplakin II
                     
                     DSPII (NP_001008844.1)
                     
                     ---
                     Assigned domains are from Choi et al., Nature Structural 
                     Biology, 2012."
                     /note="This feature has 4 segments:
                        1:  2921 ..  6094 / #008000 / N-terminus: Armadilllof...
                        2:  6095 ..  6961 / #008000 / Coiled-coiled-rod
                        3:  6962 ..  6979 / #008000 / C-terminus: Intermediat...
                        4:  8519 .. 11275 / #008000 / C-terminus: 
                     Intermediat..."
                     /translation="MSCNGGSHPRINTLGRMIRAESGPDLRYEVTSGGGGTSRMYYSRR
                     GVITDQNSDGYCQTGTMSRHQNQNTIQELLQNCSDCLMRAELIVQPELKYGDGIQLTRS
                     RELDECFAQANDQMEILDSLIREMRQMGQPCDAYQKRLLQLQEQMRALYKAISVPRVRR
                     ASSKGGGGYTCQSGSGWDEFTKHVTSECLGWMRQQRAEMDMVAWGVDLASVEQHINSHR
                     GIHNSIGDYRWQLDKIKADLREKSAIYQLEEEYENLLKASFERMDHLRQLQNIIQATSR
                     EIMWINDCEEEELLYDWSDKNTNIAQKQEAFSIRMSQLEVKEKELNKLKQESDQLVLNQ
                     HPASDKIEAYMDTLQTQWSWILQITKCIDVHLKENAAYFQFFEEAQSTEAYLKGLQDSI
                     RKKYPCDKNMPLQHLLEQIKELEKEREKILEYKRQVQNLVNKSKKIVQLKPRNPDYRSN
                     KPIILRALCDYKQDQKIVHKGDECILKDNNERSKWYVTGPGGVDMLVPSVGLIIPPPNP
                     LAVDLSCKIEQYYEAILALWNQLYINMKSLVSWHYCMIDIEKIRAMTIAKLKTMRQEDY
                     MKTIADLELHYQEFIRNSQGSEMFGDDDKRKIQSQFTDAQKHYQTLVIQLPGYPQHQTV
                     TTTEITHHGTCQDVNHNKVIETNRENDKQETWMLMELQKIRRQIEHCEGRMTLKNLPLA
                     DQGSSHHITVKINELKSVQNDSQAIAEVLNQLKDMLANFRGSEKYCYLQNEVFGLFQKL
                     ENINGVTDGYLNSLCTVRALLQAILQTEDMLKVYEARLTEEETVCLDLDKVEAYRCGLK
                     KIKNDLNLKKSLLATMKTELQKAQQIHSQTSQQYPLYDLDLGKFGEKVTQLTDRWQRID
                     KQIDFRLWDLEKQIKQLRNYRDNYQAFCKWLYDAKRRQDSLESMKFGDSNTVMRFLNEQ
                     KNLHSEISGKRDKSEEVQKIAELCANSIKDYELQLASYTSGLETLLNIPIKRTMIQSPS
                     GVILQEAADVHARYIELLTRSGDYYRFLSEMLKSLEDLKLKNTKIEVLEEELRLARDAN
                     SENCNKNKFLDQNLQKYQAECSQFKAKLASLEELKRQAELDGKSAKQNLDKCYGQIKEL
                     NEKITRLTYEIEDEKRRRKSVEDRFDQQKNDYDQLQKARQCEKENLGWQKLESEKAIKE
                     KEYEIERLRVLLQEEGTRKREYENELAKASNRIQESKNQCTQVVQERESLLVKIKVLEQ
                     DKARLQRLEDELNRAKSTLEAETRVKQRLECEKQQIQNDLNQWKTQYSRKEEAIRKIES
                     EREKSEREKNSLRSEIERLQAEIKRIEERCRRKLEDSTRETQSQLETERSRYQREIDKL
                     RQRPYGSHRETQTECEWTVDTSKLVFDGLRKKVTAMQLYECQLIDKTTLDKLLKGKKSV
                     EEVASEIQPFLRGAGSIAGASASPKEKYSLVEAKRKKLISPESTVMLLEAQAATGGIID
                     PHRNEKLTVDSAIARDLIDFDDRQQIYAAEKAITGFDDPFSGKTVSVSEAIKKNLIDRE
                     TGMRLLEAQIASGGVVDPVNSVFLPKDVALARGLIDRDLYRSLNDPRDSQKNFVDPVTK
                     KKVSYVQLKERCRIEPHTGLLLLSVQKRSMSFQGIRQPVTVTELVDSGILRPSTVNELE
                     SGQISYDEVGERIKDFLQGSSCIAGIYNETTKQKLGIYEAMKIGLVRPGTALELLEAQA
                     ATGFIVDPVSNLRLPVEEAYKRGLVGIEFKEKLLSAERAVTGYNDPETGNIISLFQAMN
                     KELIEKGHGIRLLEAQIATGGIIDPKESHRLPVDIAYKRGYFNEELSEILSDPSDDTKG
                     FFDPNTEENLTYLQLKERCIKDEETGLCLLPLKEKKKQVQTSQKNTLRKRRVVIVDPET
                     NKEMSVQEAYKKGLIDYETFKELCEQECEWEEITITGSDGSTRVVLVDRKTGSQYDIQD
                     AIDKGLVDRKFFDQYRSGSLSLTQFADMISLKNGVGTSSSMGSGVSDDVFSSSRHESVS
                     KISTISSVRNLTIRSSSFSDTLEESSPIAAIFDTENLEKISITEGIERGIVDSITGQRL
                     LEAQACTGGIIHPTTGQKLSLQDAVSQGVIDQDMATRLKPAQKAFIGFEGVKGKKKMSA
                     AEAVKEKWLPYEAGQRFLEFQYLTGGLVDPEVHGRISTEEAIRKGFIDGRAAQRLQDTS
                     SYAKILTCPKTKLKISYKDAINRSMVEDITGLRLLEAASVSSKGLPSPYNMSSAPGSRS
                     GSRSGSRSGSRSGSRSGSRRGSFDATGNSSYSYSYSFSSSSIGH"
     misc_feature    6980..6985
                     /label=dXhoI/dSalI
                     /note="color: #ff00ff"
     CDS             6986..8509
                     /codon_start=1
                     /label=F40-based tension sensor with dead YPet
                     /note="acceptor only control for tension sensor 
                     measurements:
                     F40-based tension sensor module (Austen et al., Nature Cell
                     Biology, 2015)
                     with F40 peptide that is sensitive to 1-6 pN (Grashoff et 
                     al., Nature, 2010) 
                     with YPet(short) (Y67G) mutation to disrupt chromophore 
                     formation (Zacharias and Tsien, Methods of Biochemical 
                     Analysis, Green Fluorescent Proteins, 2005)"
                     /note="color: #800080"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGGGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGSGPGGAGPGGAGPGGAGPGGAGPGGAGPGGAGPGGAGPGGAGSMVSKGEEDN
                     MAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSP
                     QFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKV
                     KLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTY
                     KAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYK"
     CDS             6986..7669
                     /codon_start=1
                     /product="short version of YPet  
                     yellow fluorescent protein optimized for FRET with CyPet 
                     (Nguyen and Daugherty, Nature Biotechnology, 2005)"
                     /label=dead YPet (short)
                     /note="mammalian codon-optimized"
                     /note="color: #808080"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGGGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAA"
     misc_feature    7184..7186
                     /label=Y67G
                     /note="dead YPet mutation"
                     /note="color: #ff0000"
     CDS             7676..7795
                     /codon_start=1
                     /label=F40
                     /note="Flagelliform linker peptide sensitive to 1-6 pN 
                     (Grashoff et al., Nature, 2010)"
                     /note="color: #ccffcc"
                     /translation="GPGGAGPGGAGPGGAGPGGAGPGGAGPGGAGPGGAGPGGA"
     CDS             7802..8509
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., Nature Biotechnology, 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /note="color: #f05078"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    8510..8517
                     /label=NotI
                     /note="NotI recognition site"
                     /note="color: #0080ff"
     misc_feature    11279..11284
                     /label=ClaI
                     /note="ClaI recognition site"
                     /note="color: #0080ff"
     primer_bind     complement(11321..11346)
                     /label=pLPCX_seq_r
                     /note="pLPCX_seq_r"
                     /note="color: blue; sequence: ACCTACAGGTGGGGTCTTTCATTCCC"
     LTR             11324..11917
                     /label=3' LTR
                     /note="3' MoMuLV LTR (long terminal repeat from Moloney 
                     murine leukemia virus)"
                     /note="color: #f0e1e1"
     protein_bind    complement(12170..12203)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: LEFT"
     protein_bind    complement(12263..12296)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: LEFT"
     rep_origin      12586
                     /label=x
                     /note="site of replication initiation"
                     /note="color: #000000"
     rep_origin      complement(12587..13175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff99"
     CDS             complement(13346..14206)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 13346 .. 14137 / #ccffcc
                        2: 14138 .. 14206 / #ccffcc / signal sequence
                     Cleavage site after base 14137"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(14207..14311)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aatgatcccc cgggctgcag gtcggccgcc
     1501 acgaccggtg ccgccaccat cccctgaccc acgcccctga cccctcacaa ggagacgacc
     1561 ttccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
     1621 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
     1681 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     1741 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     1801 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     1861 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     1921 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     1981 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     2041 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     2101 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     2161 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc
     2221 cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag gcccaccgac
     2281 tctagtaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     2341 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     2401 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     2461 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     2521 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     2581 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2641 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     2701 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     2761 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     2821 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
     2881 ggactcagat ctgggcccat aaagcttata cgaattcacc atgagctgca acggaggctc
     2941 ccacccgcgg atcaacactc tgggccgcat gatccgcgcc gagtctggcc cggacctgcg
     3001 ctacgaggtg accagcggcg gcgggggcac cagcaggatg tactattctc ggcgcggcgt
     3061 gatcaccgac cagaactcgg acggctactg tcaaaccggc acgatgtcca ggcaccagaa
     3121 ccagaacacc atccaggagc tgctgcagaa ctgctccgac tgcttgatgc gagcagagct
     3181 catcgtgcag cctgaattga agtatggaga tggaatacaa ctgactcgga gtcgagaatt
     3241 ggatgagtgt tttgcccagg ccaatgacca aatggaaatc ctcgacagct tgatcagaga
     3301 gatgcggcag atgggccagc cctgtgatgc ttaccagaaa aggcttcttc agctccaaga
     3361 gcaaatgcga gccctttata aagccatcag tgtcccacga gtccgcaggg ccagctccaa
     3421 gggtggtgga ggctacactt gtcagagtgg ctctggctgg gatgagttca ccaaacatgt
     3481 caccagtgaa tgtttggggt ggatgaggca gcaaagggcg gagatggaca tggtggcctg
     3541 gggtgtggac ctggcctcag tggagcagca cattaacagc caccggggca tccacaactc
     3601 catcggcgac tatcgctggc agctggacaa aatcaaagcc gacctgcgcg agaaatctgc
     3661 gatctaccag ttggaggagg agtatgaaaa cctgctgaaa gcgtcctttg agaggatgga
     3721 tcacctgcga cagctgcaga acatcattca ggccacgtcc agggagatca tgtggatcaa
     3781 tgactgcgag gaggaggagc tgctgtacga ctggagcgac aagaacacca acatcgctca
     3841 gaaacaggag gccttctcca tacgcatgag tcaactggaa gttaaagaaa aagagctcaa
     3901 taagctgaaa caagaaagtg accaacttgt cctcaatcag catccagctt cagacaaaat
     3961 tgaggcctat atggacactc tgcagacgca gtggagttgg attcttcaga tcaccaagtg
     4021 cattgatgtt catctgaaag aaaatgctgc ctactttcag ttttttgaag aggcgcagtc
     4081 tactgaagca tacctgaagg ggctccagga ctccatcagg aagaagtacc cctgcgacaa
     4141 gaacatgccc ctgcagcacc tgctggaaca gatcaaggag ctggagaaag aacgagagaa
     4201 aatccttgaa tacaagcgtc aggtgcagaa cttggtaaac aagtctaaga agattgtaca
     4261 gctgaagcct cgtaacccag actacagaag caataaaccc attattctca gagctctctg
     4321 tgactacaaa caagatcaga aaatcgtgca taagggggat gagtgtatcc tgaaggacaa
     4381 caacgagcgc agcaagtggt acgtgacggg cccgggaggc gttgacatgc ttgttccctc
     4441 tgtggggctg atcatccctc ctccgaaccc actggccgtg gacctctctt gcaagattga
     4501 gcagtactac gaagccatct tggctctgtg gaaccagctc tacatcaaca tgaagagcct
     4561 ggtgtcctgg cactactgca tgattgacat agagaagatc agggccatga caatcgccaa
     4621 gctgaaaaca atgcggcagg aagattacat gaagacgata gccgaccttg agttacatta
     4681 ccaagagttc atcagaaata gccaaggctc agagatgttt ggagatgatg acaagcggaa
     4741 aatacagtct cagttcaccg atgcccagaa gcattaccag accctggtca ttcagctccc
     4801 tggctatccc cagcaccaga cagtgaccac aactgaaatc actcatcatg gaacctgcca
     4861 agatgtcaac cataataaag taattgaaac caacagagaa aatgacaagc aagaaacatg
     4921 gatgctgatg gagctgcaga agattcgcag gcagatagag cactgcgagg gcaggatgac
     4981 tctcaaaaac ctccctctag cagaccaggg gtcttctcac cacatcacag tgaaaattaa
     5041 cgagcttaag agtgtgcaga atgattcaca agcaattgct gaggttctca accagcttaa
     5101 agatatgctt gccaacttca gaggttctga aaagtactgc tatttacaga atgaagtatt
     5161 tggactattt cagaaactgg aaaatatcaa tggtgttaca gatggctact taaatagctt
     5221 atgcacagta agggcactgc tccaggctat tctccaaaca gaagacatgt taaaggttta
     5281 tgaagccagg ctcactgagg aggaaactgt ctgcctggac ctggataaag tggaagctta
     5341 ccgctgtgga ctgaagaaaa taaaaaatga cttgaacttg aagaagtcgt tgttggccac
     5401 tatgaagaca gaactacaga aagcccagca gatccactct cagacttcac agcagtatcc
     5461 actttatgat ctggacttgg gcaagttcgg tgaaaaagtc acacagctga cagaccgctg
     5521 gcaaaggata gataaacaga tcgactttag attatgggac ctggagaaac aaatcaagca
     5581 attgaggaat tatcgtgata actatcaggc tttctgcaag tggctctatg atgctaaacg
     5641 ccgccaggat tccttagaat ccatgaaatt tggagattcc aacacagtca tgcggttttt
     5701 gaatgagcag aagaacttgc acagtgaaat atctggcaaa cgagacaaat cagaggaagt
     5761 acaaaaaatt gctgaacttt gcgccaattc aattaaggat tatgagctcc agctggcctc
     5821 atacacctca ggactggaaa ctctgctgaa catacctatc aagaggacca tgattcagtc
     5881 cccttctggg gtgattctgc aagaggctgc agatgttcat gctcggtaca ttgaactact
     5941 tacaagatct ggagactatt acaggttctt aagtgagatg ctgaagagtt tggaagatct
     6001 gaagctgaaa aataccaaga tcgaagtttt ggaagaggag ctcagactgg cccgagatgc
     6061 caactcggaa aactgtaata agaacaaatt cctggatcag aacctgcaga aataccaggc
     6121 agagtgttcc cagttcaaag cgaagcttgc gagcctggag gagctgaaga gacaggctga
     6181 gctggatggg aagtcggcta agcaaaatct agacaagtgc tacggccaaa taaaagaact
     6241 caatgagaag atcacccgac tgacttatga gattgaagat gaaaagagaa gaagaaaatc
     6301 tgtggaagac agatttgacc aacagaagaa tgactatgac caactgcaga aagcaaggca
     6361 atgtgaaaag gagaaccttg gttggcagaa attagagtct gagaaagcca tcaaggagaa
     6421 ggagtacgag attgaaaggt tgagggttct actgcaggaa gaaggcaccc ggaagagaga
     6481 atatgaaaat gagctggcaa aggcatctaa taggattcag gaatcaaaga atcagtgtac
     6541 tcaggtggta caggaaagag agagccttct ggtgaaaatc aaagtcctgg agcaagacaa
     6601 ggcaaggctg cagaggctgg aggatgagct gaatcgtgca aaatcaactc tagaggcaga
     6661 aaccagggtg aaacagcgcc tggagtgtga gaaacagcaa attcagaatg acctgaatca
     6721 gtggaagact caatattccc gcaaggagga ggctattagg aagatagaat cggaaagaga
     6781 aaagagtgag agagagaaga acagtcttag gagtgagatc gaaagactcc aagcagagat
     6841 caagagaatt gaagagaggt gcaggcgtaa gctggaggat tctaccaggg agacacagtc
     6901 acagttagaa acagaacgct cccgatatca gagggagatt gataaactca gacagcgccc
     6961 atatgggtcc catcgagagg tcgagatggt gagcaaaggc gaagagctgt tcaccggcgt
     7021 ggtgcccatc ctggtggagc tggacggcga cgtgaacggc cacaagttca gcgtgagcgg
     7081 cgagggcgag ggcgacgcca cctacggcaa gctgaccctg aagctgctgt gcaccaccgg
     7141 caagctgccc gtgccctggc ccaccctggt gaccaccctg ggcggcggcg tgcagtgctt
     7201 cgcccggtac cccgaccaca tgaagcagca cgacttcttc aagagcgcca tgcccgaggg
     7261 ctacgtgcag gagcggacca tcttcttcaa ggacgacggc aactacaaga cccgggccga
     7321 ggtgaagttc gagggcgaca ccctggtgaa ccggatcgag ctgaagggca tcgacttcaa
     7381 ggaggacggc aacatcctgg gccacaagct ggagtacaac tacaacagcc acaacgtgta
     7441 catcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc ggcacaacat
     7501 cgaggacggc ggcgtgcagc tggccgacca ctaccagcag aacaccccca tcggcgacgg
     7561 ccccgtgctg ctgcccgaca accactacct gagctaccag agcgccctgt tcaaggaccc
     7621 caacgagaag cgggaccaca tggtgctgct ggagttcctg accgccgccg gatccgggcc
     7681 aggtggagca gggccaggtg gtgcagggcc aggtggtgca gggccaggtg gtgcagggcc
     7741 cggtggtgca ggtccaggtg gtgcaggtcc aggtggtgct ggtccaggtg gtgctggatc
     7801 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     7861 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     7921 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     7981 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     8041 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     8101 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     8161 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     8221 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     8281 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     8341 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     8401 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     8461 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg cggccgctac
     8521 ccagactgag tgtgagtgga ccgttgacac ctccaagctg gtgtttgatg ggctgaggaa
     8581 gaaggtgaca gcaatgcagc tctatgagtg tcagctgatc gacaaaacaa ccttggacaa
     8641 actattgaag gggaagaagt cagtggaaga agttgcttct gaaatccagc cattccttcg
     8701 gggtgcagga tctatcgctg gagcatctgc ttctcctaag gaaaaatact ctttggtaga
     8761 ggccaagaga aagaaattaa tcagcccaga atccacagtc atgcttctgg aggcccaggc
     8821 agctacaggt ggtataattg atccccatcg gaatgagaag ctgactgtgg acagtgccat
     8881 agctcgggac ctcattgact tcgatgaccg tcagcagata tatgcagcag aaaaagctat
     8941 cactggtttt gatgatccat tttcaggcaa gacagtatct gtttcagaag ccatcaagaa
     9001 aaatttgatt gatagagaaa ccggaatgcg cctgctggaa gcccagattg cttcaggggg
     9061 tgtagtagac cctgtgaaca gtgtcttttt gccaaaagat gtcgccttgg cccgggggct
     9121 gattgataga gatttgtatc gttccctgaa tgatccccga gatagtcaga aaaactttgt
     9181 ggatccagtc accaaaaaga aggtcagtta cgtgcagctg aaggaacggt gcagaatcga
     9241 accacatact ggtctgctct tgctttcagt acagaagaga agcatgtcct tccaaggaat
     9301 cagacaacct gtgaccgtca ctgagctagt agattctggt atattgagac cgtccactgt
     9361 caatgaactg gaatctggtc agatttctta tgacgaggtt ggtgagagaa ttaaggactt
     9421 cctccagggt tcaagctgca tagcaggcat atacaatgag accacaaaac agaagcttgg
     9481 catttatgag gccatgaaaa ttggcttagt ccgacctggt actgctctgg agttgctgga
     9541 agcccaagca gctactggct ttatagtgga tcctgttagc aacttgaggt taccagtgga
     9601 ggaagcctac aagagaggtc tggtgggcat tgagttcaaa gagaagctcc tgtctgcaga
     9661 acgagctgtc actgggtata atgatcctga aacaggaaac atcatctctt tgttccaagc
     9721 catgaataag gaactcatcg aaaagggcca cggtattcgc ttattagaag cacagatcgc
     9781 aaccgggggg atcattgacc caaaggagag ccatcgttta ccagttgaca tagcatataa
     9841 gaggggctat ttcaatgagg aactcagtga gattctctca gatccaagtg atgataccaa
     9901 aggatttttt gaccccaaca ctgaagaaaa tcttacctat ctgcaactaa aagaaagatg
     9961 cattaaggat gaggaaacag ggctctgtct tctgcctctg aaagaaaaga agaaacaggt
    10021 gcagacatca caaaagaata ccctcaggaa gcgtagagtg gtcatagttg acccagaaac
    10081 caataaagaa atgtctgttc aggaggccta caagaagggc ctaattgatt atgaaacctt
    10141 caaagaactg tgtgagcagg aatgtgaatg ggaagaaata accatcacgg gatcagatgg
    10201 ctccaccagg gtggtcctgg tagatagaaa gacaggcagt cagtatgata ttcaagatgc
    10261 tattgacaag ggccttgttg acaggaagtt ctttgatcag taccgatccg gcagcctcag
    10321 cctcactcaa tttgctgaca tgatctcctt gaaaaatggt gtcggcacca gcagcagcat
    10381 gggcagtggt gtcagcgatg atgtttttag cagctcccga catgaatcag taagtaagat
    10441 ttccaccata tccagcgtca ggaatttaac cataaggagc agctcttttt cagacaccct
    10501 ggaagaatcg agccccattg cagccatctt tgacacagaa aacctggaga aaatctccat
    10561 tacagaaggt atagagcggg gcatcgttga cagcatcacg ggtcagaggc ttctggaggc
    10621 tcaggcctgc acaggtggca tcatccaccc aaccacgggc cagaagctgt cacttcagga
    10681 cgcagtctcc cagggtgtga ttgaccaaga catggccacc aggctgaagc ctgctcagaa
    10741 agccttcata ggcttcgagg gtgtgaaggg aaagaagaag atgtcagcag cagaggcagt
    10801 gaaagaaaaa tggctcccgt atgaggctgg ccagcgcttc ctggagttcc agtacctcac
    10861 gggaggtctt gttgacccgg aagtgcatgg gaggataagc accgaagaag ccatccggaa
    10921 ggggttcata gatggccgcg ccgcacagag gctgcaagac accagcagct atgccaaaat
    10981 cctgacctgc cccaaaacca aattaaaaat atcctataag gatgccataa atcgctccat
    11041 ggtagaagat atcactgggc tgcgccttct ggaagccgcc tccgtgtcgt ccaagggctt
    11101 acccagccct tacaacatgt cttcggctcc ggggtcccgc tccggctccc gctcgggatc
    11161 tcgctccgga tctcgctccg ggtcccgcag tgggtcccgg agaggaagct ttgacgccac
    11221 aggcaattct tcctactctt attcctactc atttagcagt agttctattg ggcactaaat
    11281 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
    11341 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
    11401 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
    11461 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
    11521 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
    11581 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
    11641 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
    11701 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
    11761 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
    11821 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
    11881 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
    11941 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt
    12001 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
    12061 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
    12121 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtactga taacttcgta
    12181 taatgtatgc tatacgaagt tattaggtct gaagaggagt ttacgtccag ccaagctagg
    12241 gccgcgatcc ggaaccctta atataacttc gtataatgta tgctatacga agttatcagt
    12301 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
    12361 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
    12421 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
    12481 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
    12541 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
    12601 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
    12661 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
    12721 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
    12781 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
    12841 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
    12901 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
    12961 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
    13021 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
    13081 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
    13141 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
    13201 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
    13261 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
    13321 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
    13381 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
    13441 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
    13501 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
    13561 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
    13621 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
    13681 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
    13741 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
    13801 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
    13861 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
    13921 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
    13981 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
    14041 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
    14101 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
    14161 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
    14221 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
    14281 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
    14341 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
    14401 ttcgtcttca agaattgcta gcaattgcta gcaattgcta gcaattcata ccagatcacc
    14461 gaaaactgtc ctccaaatgt gtccccctca cactcccaaa ttcgcgggct tctgcctctt
    14521 agaccactct accctattcc ccacactcac cggagccaaa gccgcggccc ttccgtttct
    14581 ttgct
//
