pEBTet_NAD-Snifit-mito

LOCUS       pEBTet_NAD-Snifit-mito               14157 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Plasmid encoding the mitochondrial localization of the FRET 
            semi-synthetic biosensor measuring NAD+ concentration in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    pEBTet_NAD-Snifit-mito
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14157)
  AUTHORS   Sallin O, Reymond L, Gondrand C, Raith F, Koch B, Johnsson K
  TITLE     Semisynthetic biosensors for mapping cellular concentrations of 
            nicotinamide adenine dinucleotides.
  JOURNAL   Elife. 2018 May 29;7. pii: 32638. doi: 10.7554/eLife.32638.
  PUBMED    29809136
REFERENCE   2  (bases 1 to 14157)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14157
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             12..86
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             99..170
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             189..260
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             273..344
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1167..2051
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(2782..2799)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2788..2899
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3440..5229
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     primer_bind     complement(7893..7911)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7979..8083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8084..8944
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8302..8321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9115..9703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9604..9623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(9892..9912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9910..10207
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     10139..10158
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             10284..10883
                     /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(10284..10303)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10780..10800
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    10974..11108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11246..11265)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        11542..11803
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     intron          11920..12492
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     12500..12519
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     12547..12566
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        12547..12565
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             12576..13193
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
     primer_bind     complement(12649..12668)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     polyA_signal    13238..13359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        13414..13793
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        13794..13997
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     13948..13968
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    13999..14017
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14020..14038
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     14048..14072
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 agcttaccac catgtccgtc ctgacgccgc tgctgctgcg gggcttgaca ggctcggccc
       61 ggcggctccc agtgccgcgc gccaagatcc attcgttgtc cgtcctgacg ccgctgctgc
      121 tgcggggctt gacaggctcg gcccggcggc tcccagtgcc gcgcgccaag atccattcgt
      181 tgggaggcag cgtgctcacc ccactcctgc tgcggggctt gacaggctcg gcccggcggc
      241 tcccagtgcc gcgcgctaaa attcacagcc tctccgtcct gacgccgctg ctgctgcggg
      301 gcttgacagg ctcggcccgg cggctcccag tgccgcgcgc caagatccat tcgttgggag
      361 gcagcggagg cagcgagggc gggctggggc gtgctgtgtg cttgctgacc ggggcctccc
      421 gcggcttcgg ccggacgctg gccccgctcc tggcctcgct gctgtcgccc ggctccgtgc
      481 ttgtccttag cgactggaac gacgaggcac tgcgccagct ggaggccgag ctgggcgccg
      541 agcggtctgg cctgcgcgtg gtgcgggtgc ccgccgacct gggcgccgag gccggcttgc
      601 agcagctgct cggcgccctg cgcgagctcc cccggcccaa ggggctgcag cgactgctgc
      661 ttatcaacaa cgcgggctct cttggggatg tgtccaaagg cttcgtggac ctgagtgact
      721 ccactcaagt gaacaactac tgggcactga acttgacctc catgctctgc ctgacttcca
      781 gcgtcctgaa ggccttcccg gacagtcctg gcctcaacag aaccgtggtt aacatctcgt
      841 ccctctgtgc cctgcaacct ttcaaaggct gggcgctgta ctgtgcagga aaggctgctc
      901 gtgatatgct gttccaggtc ctggcgctgg aggaacctaa tgtgagggtg ctgaactatg
      961 ccccaggtcc tctggacaca gacatgcagc agttggcccg ggagacctcc gtggacccag
     1021 acatgcgaaa agggctgcag gagctgaagg caaaggggaa gctggtggat tgcaaggtgt
     1081 cagcccagaa actgctgagc ttactggaaa aggacgagtt caagtctgga gcccacgtgg
     1141 acttctatga caaagaattc ggatccgaaa tcggtactgg ctttccattc gacccccatt
     1201 atgtggaagt cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc
     1261 ctgtgctgtt cctgcacggt aacccgacct cctcctacgt gtggcgcaac atcatcccgc
     1321 atgttgcacc gacccatcgc tgcattgctc cagacctgat cggtatgggc aaatccgaca
     1381 aaccagacct gggttatttc ttcgacgacc acgtccgctt catggatgcc ttcatcgaag
     1441 ccctgggtct ggaagaggtc gtcctggtca ttcacgactg gggctccgct ctgggtttcc
     1501 actgggccaa gcgcaatcca gagcgcgtca aaggtattgc atttatggag ttcatccgcc
     1561 ctatcccgac ctgggacgaa tggccagaat ttgcccgcga gaccttccag gccttccgca
     1621 ccaccgacgt cggccgcaag ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc
     1681 cgatgggtgt cgtccgcccg ctgactgaag tcgagatgga ccattaccgc gagccgttcc
     1741 tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca atcgccggtg
     1801 agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac cagtcccctg
     1861 tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc gaagccgctc
     1921 gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt ctgaatctgc
     1981 tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctg tctactctgg
     2041 agatttccgg tagcggccgc cctcctcctc cgccaccgcc accaccgccg ccgccgccac
     2101 caccacctcc tccgcctcct ccaccaccac cgccaccgcc tccaccacct ggcggccgct
     2161 ctcggtctct cgagatggat aaagactgcg aaatgaaacg cactaccctg gacagcccgc
     2221 tgggtaaact ggaactgagc ggttgcgagc agggcctgca cgaaatcatt ttcctgggca
     2281 aaggtacgag cgctgcggat gcggtcgaag taccggctcc ggctgcggtt ctgggtggcc
     2341 cggagccgct gatgcaggct accgcatggc tgaacgcata ttttcaccag ccagaggcga
     2401 ttgaagagtt ccctgtaccg gccctgcatc acccggtatt tcagcaggaa tcctttaccc
     2461 gtcaggtact gtggaaactg ctgaaagtag tgaaattcgg cgaggtgatc tcctattccc
     2521 atctggcggc gctggctggc aacccagcgg caactgcggc ggtgaaaacc gctctgtctg
     2581 gtaatccggt gccaattctg atcccgtgcc accgtgtggt tcagggcgac ctggacgtag
     2641 gtggctacga aggcggcctg gctgtgaaag aatggctgct ggcgcacgaa ggtcaccgtc
     2701 tgggtaaacc aggcctgggc taaagctagc ggcgcgcccg gccgctcgag tctagagggc
     2761 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2821 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2881 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2941 tggggcagga cagcaagggg gaggattggg aagacaatag caggctagag gatcgatccc
     3001 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     3061 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     3121 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     3181 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     3241 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     3301 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     3361 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     3421 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     3481 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     3541 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     3601 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     3661 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     3721 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     3781 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     3841 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     3901 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     3961 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     4021 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     4081 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     4141 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     4201 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     4261 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     4321 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     4381 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     4441 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     4501 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     4561 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     4621 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     4681 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4741 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     4801 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     4861 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4921 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4981 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     5041 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     5101 gtagcatatg ctatcctcat gcatatacag tcagcatatg atacccagta gtagagtggg
     5161 agtgctatcc tttgcatatg ccgccacctc ccaagggggc gtgaattttc gctgcttgtc
     5221 cttttcctgc tggttgctcc cattcttagg tgaatttaag gaggccaggc taaagccgtc
     5281 gcatgtctga ttgctcacca ggtaaatgtc gctaatgttt tccaacgcga gaaggtgttg
     5341 agcgcggagc tgagtgacgt gacaacatgg gtatgcccaa ttgccccatg ttgggaggac
     5401 gaaaatggtg acaagacaga tggccagaaa tacaccaaca gcacgcatga tgtctactgg
     5461 ggatttattc tttagtgcgg gggaatacac ggcttttaat acgattgagg gcgtctccta
     5521 acaagttaca tcactcctgc ccttcctcac cctcatctcc atcacctcct tcatctccgt
     5581 catctccgtc atcaccctcc gcggcagccc cttccaccat aggtggaaac cagggaggca
     5641 aatctactcc atcgtcaaag ctgcacacag tcaccctgat attgcaggta ggagcgggct
     5701 ttgtcataac aaggtcctta atcgcatcct tcaaaacctc agcaaatata tgagtttgta
     5761 aaaagaccat gaaataacag acaatggact cccttagcgg gccaggttgt gggccgggtc
     5821 caggggccat tccaaagggg agacgactca atggtgtaag acgacattgt ggaatagcaa
     5881 gggcagttcc tcgccttagg ttgtaaaggg aggtcttact acctccatat acgaacacac
     5941 cggcgaccca agttccttcg tcggtagtcc tttctacgtg actcctagcc aggagagctc
     6001 ttaaaccttc tgcaatgttc tcaaatttcg ggttggaacc tccttgacca cgatgctttt
     6061 ccaaaccacc ctcctttttt gcgccctgcc tccatcaccc tgaccccggg gtccagtgct
     6121 tgggccttct cctgggtcat ctgcggggcc ctgctctatc gctcccgggg gcacgtcagg
     6181 ctcaccatct gggccacctt cttggtggta ttcaaaataa tcggcttccc ctacagggtg
     6241 gaaaaatggc cttctacctg gagggggcct gcgcggtgga gacccggatg atgatgactg
     6301 actactggga ctcctgggcc tcttttctcc acgtccacga cctctccccc tggctctttc
     6361 acgacttccc cccctggctc tttcacgtcc tctaccccgg cggcctccac tacctcctcg
     6421 accccggcct ccactacctc ctcgaccccg gcctccactg cctcctcgac cccggcctcc
     6481 acctcctgct cctgcccctc ctgctcctgc ccctcctcct gctcctgccc ctcctgcccc
     6541 tcctgctcct gcccctcctg cccctcctgc tcctgcccct cctgcccctc ctgctcctgc
     6601 ccctcctgcc cctcctcctg ctcctgcccc tcctgcccct cctcctgctc ctgcccctcc
     6661 tgcccctcct gctcctgccc ctcctgcccc tcctgctcct gcccctcctg cccctcctgc
     6721 tcctgcccct cctgctcctg cccctcctgc tcctgcccct cctgctcctg cccctcctgc
     6781 ccctcctgcc cctcctcctg ctcctgcccc tcctgctcct gcccctcctg cccctcctgc
     6841 ccctcctgct cctgcccctc ctcctgctcc tgcccctcct gcccctcctg cccctcctcc
     6901 tgctcctgcc cctcctgccc ctcctcctgc tcctgcccct cctcctgctc ctgcccctcc
     6961 tgcccctcct gcccctcctc ctgctcctgc ccctcctgcc cctcctcctg ctcctgcccc
     7021 tcctcctgct cctgcccctc ctgcccctcc tgcccctcct cctgctcctg cccctcctcc
     7081 tgctcctgcc cctcctgccc ctcctgcccc tcctgcccct cctcctgctc ctgcccctcc
     7141 tcctgctcct gcccctcctg ctcctgcccc tcccgctcct gctcctgctc ctgttccacc
     7201 gtgggtccct ttgcagccaa tgcaacttgg acgtttttgg ggtctccgga caccatctct
     7261 atgtcttggc cctgatcctg agccgcccgg ggctcctggt cttccgcctc ctcgtcctcg
     7321 tcctcttccc cgtcctcgtc catggttatc accccctctt ctttgaggtc cactgccgcc
     7381 ggagccttct ggtccagatg tgtctccctt ctctcctagg ccatttccag gtcctgtacc
     7441 tggcccctcg tcagacatga ttcacactaa aagagatcaa tagacatctt tattagacga
     7501 cgctcagtga atacagggag tgcagactcc tgccccctcc aacagccccc ccaccctcat
     7561 ccccttcatg gtcgctgtca gacagatcca ggtctgaaaa ttccccatcc tccgaaccat
     7621 cctcgtcctc atcaccaatt actcgcagcc cggaaaactc ccgctgaaca tcctcaagat
     7681 ttgcgtcctg agcctcaagc caggcctcaa attcctcgtc cccctttttg ctggacggta
     7741 gggatgggga ttctcgggac ccctcctctt cctcttcaag gtcaccagac agagatgcta
     7801 ctggggcaac ggaagaaaag ctgggtgcgg cctgtgagga tcagcttatc gatgataagc
     7861 tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     7921 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7981 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     8041 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     8101 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     8161 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     8221 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     8281 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     8341 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     8401 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     8461 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     8521 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     8581 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     8641 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     8701 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8761 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     8821 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8881 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8941 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     9001 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     9061 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     9121 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     9181 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     9241 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     9301 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     9361 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     9421 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     9481 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     9541 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     9601 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     9661 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     9721 ctttttacgg ttcctggcct tttgctggcc ttgaagctgt ccctgatggt cgtcatctac
     9781 ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat
     9841 cataatgggg aaggccatcc agcctcgcgt cgaactagat gatccggctg tggaatgtgt
     9901 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     9961 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
    10021 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
    10081 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
    10141 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
    10201 tttttggagg gtgaccgcca cgaccggtgc cgccaccatc ccctgaccca cgcccctgac
    10261 ccctcacaag gagacgacct tccatgaccg agtacaagcc cacggtgcgc ctcgccaccc
    10321 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
    10381 cgcgccacac cgtcgacccc gaccgccaca tcgaacgcgt caccgagctg caagaactct
    10441 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
    10501 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
    10561 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
    10621 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
    10681 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
    10741 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
    10801 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
    10861 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga
    10921 gcgcacgacc cggtccgacg cggcccgacg ggtccgaggg gggtcgacct cgaaacttgt
    10981 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
    11041 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
    11101 tctggatctg atccgaaccc cttcctcgac caattctcat gtttgacagc ttatcatcgc
    11161 agatccgggc aacgttgttg ccattgctcg accaattctc atgtttgaca gcttatcatc
    11221 gcagatccgt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
    11281 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
    11341 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
    11401 cgctgcttcg cgatgtacgg gccagatata cgcgtatctg aggggactag ggtgtgttta
    11461 ggcgaaaagc ggggcttcgg ttgtacgcgg ttaggagtcc cctcaggata tagtagtttc
    11521 gcttttgcat agggaggggg aaatgtagtc ttatgcaata ctcttgtagt cttgcaacat
    11581 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
    11641 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
    11701 gattggacga accactgaat tccgcattgc agagatattg tatttaagtg cctagctcga
    11761 tacaataaac gccatttgac cattcaccac attggtgtgc acctccaagc tggccgggga
    11821 tcctctaggg cctctgagct attccagaag tagtgaagag gcttttttgg aggcctaggc
    11881 ttttgcaaaa agctccggat cgatcctgag aacttcaggg tgagtttggg gacccttgat
    11941 tgttctttct ttttcgctat tgtaaaattc atgttatatg gagggggcaa agttttcagg
    12001 gtgttgttta gaatgggaag atgtcccttg tatcaccatg gaccctcatg ataattttgt
    12061 ttctttcact ttctactctg ttgacaacca ttgtctcctc ttattttctt ttcattttct
    12121 gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt cacttttgtt
    12181 tatttgtcag attgtaagta ctttctctaa tcactttttt ttcaaggcaa tcagggtata
    12241 ttatattgta cttcagcaca gttttagaga acaattgtta taattaaatg ataaggtaga
    12301 atatttctgc atataaattc tggctggcgt ggaaatattc ttattggtag aaacaactac
    12361 atcctggtca tcatcctgcc tttctcttta tggttacaat gatatacact gtttgagatg
    12421 aggataaaat actctgagtc caaaccgggc ccctctgcta accatgttca tgccttcttc
    12481 tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa
    12541 gaattgtaat acgactcact atagggcgaa ttgatatgtc tagattagat aaaagtaaag
    12601 tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta
    12661 aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc
    12721 gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc
    12781 ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg
    12841 ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac
    12901 agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga
    12961 atgcattata tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc
    13021 aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat
    13081 tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg
    13141 gccttgaatt gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtccgcgt
    13201 acagcggatc ccgggaattc agatcttatt aaagcagaac ttgtttattg cagcttataa
    13261 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
    13321 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggt cgaatccatg
    13381 gcgcagaact ggtaggtatg gaagatccgc gttgacattg attattgact agttattaat
    13441 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
    13501 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
    13561 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
    13621 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
    13681 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
    13741 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
    13801 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
    13861 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
    13921 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
    13981 tctatataag cagagctctc cctatcagtg atagagatct ccctatcagt gatagagatc
    14041 gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
    14101 acctccatag aagacaccgg gaccgatcca gcctccggac tctagcgttt aaactta
//