pEBTet_NADP-Snifit-nucl

LOCUS       pEBTet_NADP-Snifit-nucl               13899 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Plasmid encoding the nuclear localization of the FRET semi-synthetic
            biosensor measuring the ratio NADPH/NADP+ in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pEBTet_NADP-Snifit-nucl
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13899)
  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 13899)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13899
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             807..1691
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             2373..2393
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2397..2417
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2524..2541)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2530..2641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3182..4971
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     primer_bind     complement(7635..7653)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7721..7825
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7826..8686
                     /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(8044..8063)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8857..9445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9346..9365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(9634..9654)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9652..9949
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     9881..9900
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             10026..10625
                     /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(10026..10045)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10522..10542
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    10716..10850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10988..11007)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        11284..11545
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     intron          11662..12234
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     12242..12261
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     12289..12308
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        12289..12307
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             12318..12935
                     /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(12391..12410)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     polyA_signal    12980..13101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        13156..13535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        13536..13739
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     13690..13710
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    13741..13759
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13762..13780
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     13790..13814
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 agcttaccac catggagggc gggctggggc gtgctgtgtg cttgctgacc ggggcctccc
       61 gcggcttcgg ccggacgctg gccccgctcc tggcctcgct gctgtcgccc ggctccgtgc
      121 ttgtccttag cgcccgcaac gacgaggcac tgcgccagct ggaggccgag ctgggcgccg
      181 agcggtctgg cctgcgcgtg gtgcgggtgc ccgccgacct gggcgccgag gccggcttgc
      241 agcagctgct cggcgccctg cgcgagctcc cccggcccaa ggggctgcag cgactgctgc
      301 ttatcaacaa cgcgggctct cttggggatg tgtccaaagg cttcgtggac ctgagtgact
      361 ccactcaagt gaacaactac tgggcactga acttgacctc catgctctgc ctgacttcca
      421 gcgtcctgaa ggccttcccg gacagtcctg gcctcaacag aaccgtggtt aacatctcgt
      481 ccctctgtgc cctgcaacct ttcaaaggct gggcgctgta ctgtgcagga aaggctgctc
      541 gtgatatgct gttccaggtc ctggcgctgg aggaacctaa tgtgagggtg ctgaactatg
      601 ccccaggtcc tctggacaca gacatgcagc agttggcccg ggagacctcc gtggacccag
      661 acatgcgaaa agggctgcag gagctgaagg caaaggggaa gctggtggat tgcaaggtgt
      721 cagcccagaa actgctgagc ttactggaaa aggacgagtt caagtctgga gcccacgtgg
      781 acttctatga caaagaattc ggatccgaaa tcggtactgg ctttccattc gacccccatt
      841 atgtggaagt cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc
      901 ctgtgctgtt cctgcacggt aacccgacct cctcctacgt gtggcgcaac atcatcccgc
      961 atgttgcacc gacccatcgc tgcattgctc cagacctgat cggtatgggc aaatccgaca
     1021 aaccagacct gggttatttc ttcgacgacc acgtccgctt catggatgcc ttcatcgaag
     1081 ccctgggtct ggaagaggtc gtcctggtca ttcacgactg gggctccgct ctgggtttcc
     1141 actgggccaa gcgcaatcca gagcgcgtca aaggtattgc atttatggag ttcatccgcc
     1201 ctatcccgac ctgggacgaa tggccagaat ttgcccgcga gaccttccag gccttccgca
     1261 ccaccgacgt cggccgcaag ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc
     1321 cgatgggtgt cgtccgcccg ctgactgaag tcgagatgga ccattaccgc gagccgttcc
     1381 tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca atcgccggtg
     1441 agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac cagtcccctg
     1501 tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc gaagccgctc
     1561 gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt ctgaatctgc
     1621 tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctg tctactctgg
     1681 agatttccgg tagcggccgc cctcctcctc cgccaccgcc accaccgccg ccgccgccac
     1741 caccacctcc tccgcctcct ccaccaccac cgccaccgcc tccaccacct ggcggccgct
     1801 ctcggtctct cgagatggat aaagactgcg aaatgaaacg cactaccctg gacagcccgc
     1861 tgggtaaact ggaactgagc ggttgcgagc agggcctgca cgaaatcatt ttcctgggca
     1921 aaggtacgag cgctgcggat gcggtcgaag taccggctcc ggctgcggtt ctgggtggcc
     1981 cggagccgct gatgcaggct accgcatggc tgaacgcata ttttcaccag ccagaggcga
     2041 ttgaagagtt ccctgtaccg gccctgcatc acccggtatt tcagcaggaa tcctttaccc
     2101 gtcaggtact gtggaaactg ctgaaagtag tgaaattcgg cgaggtgatc tcctattccc
     2161 atctggcggc gctggctggc aacccagcgg caactgcggc ggtgaaaacc gctctgtctg
     2221 gtaatccggt gccaattctg atcccgtgcc accgtgtggt tcagggcgac ctggacgtag
     2281 gtggctacga aggcggcctg gctgtgaaag aatggctgct ggcgcacgaa ggtcaccgtc
     2341 tgggtaaacc aggcctgggc ggagctccgg atccaaaaaa gaagagaaag gtagatccaa
     2401 aaaagaagag aaaggtagat ccaaaaaaga agagaaagga gctccgggct tctcctcaat
     2461 aatgaagcta gcggcgcgcc cggccgctcg agtctagagg gcccgtttaa acccgctgat
     2521 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2581 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2641 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2701 gggaggattg ggaagacaat agcaggctag aggatcgatc cccgccccgg acgaactaaa
     2761 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     2821 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     2881 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     2941 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     3001 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     3061 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     3121 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     3181 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     3241 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     3301 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     3361 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     3421 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     3481 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     3541 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     3601 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     3661 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     3721 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     3781 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     3841 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     3901 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     3961 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     4021 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     4081 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     4141 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     4201 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     4261 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     4321 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4381 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4441 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4501 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     4561 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     4621 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     4681 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4741 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4801 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     4861 atgcatatac agtcagcata tgatacccag tagtagagtg ggagtgctat cctttgcata
     4921 tgccgccacc tcccaagggg gcgtgaattt tcgctgcttg tccttttcct gctggttgct
     4981 cccattctta ggtgaattta aggaggccag gctaaagccg tcgcatgtct gattgctcac
     5041 caggtaaatg tcgctaatgt tttccaacgc gagaaggtgt tgagcgcgga gctgagtgac
     5101 gtgacaacat gggtatgccc aattgcccca tgttgggagg acgaaaatgg tgacaagaca
     5161 gatggccaga aatacaccaa cagcacgcat gatgtctact ggggatttat tctttagtgc
     5221 gggggaatac acggctttta atacgattga gggcgtctcc taacaagtta catcactcct
     5281 gcccttcctc accctcatct ccatcacctc cttcatctcc gtcatctccg tcatcaccct
     5341 ccgcggcagc cccttccacc ataggtggaa accagggagg caaatctact ccatcgtcaa
     5401 agctgcacac agtcaccctg atattgcagg taggagcggg ctttgtcata acaaggtcct
     5461 taatcgcatc cttcaaaacc tcagcaaata tatgagtttg taaaaagacc atgaaataac
     5521 agacaatgga ctcccttagc gggccaggtt gtgggccggg tccaggggcc attccaaagg
     5581 ggagacgact caatggtgta agacgacatt gtggaatagc aagggcagtt cctcgcctta
     5641 ggttgtaaag ggaggtctta ctacctccat atacgaacac accggcgacc caagttcctt
     5701 cgtcggtagt cctttctacg tgactcctag ccaggagagc tcttaaacct tctgcaatgt
     5761 tctcaaattt cgggttggaa cctccttgac cacgatgctt ttccaaacca ccctcctttt
     5821 ttgcgccctg cctccatcac cctgaccccg gggtccagtg cttgggcctt ctcctgggtc
     5881 atctgcgggg ccctgctcta tcgctcccgg gggcacgtca ggctcaccat ctgggccacc
     5941 ttcttggtgg tattcaaaat aatcggcttc ccctacaggg tggaaaaatg gccttctacc
     6001 tggagggggc ctgcgcggtg gagacccgga tgatgatgac tgactactgg gactcctggg
     6061 cctcttttct ccacgtccac gacctctccc cctggctctt tcacgacttc cccccctggc
     6121 tctttcacgt cctctacccc ggcggcctcc actacctcct cgaccccggc ctccactacc
     6181 tcctcgaccc cggcctccac tgcctcctcg accccggcct ccacctcctg ctcctgcccc
     6241 tcctgctcct gcccctcctc ctgctcctgc ccctcctgcc cctcctgctc ctgcccctcc
     6301 tgcccctcct gctcctgccc ctcctgcccc tcctgctcct gcccctcctg cccctcctcc
     6361 tgctcctgcc cctcctgccc ctcctcctgc tcctgcccct cctgcccctc ctgctcctgc
     6421 ccctcctgcc cctcctgctc ctgcccctcc tgcccctcct gctcctgccc ctcctgctcc
     6481 tgcccctcct gctcctgccc ctcctgctcc tgcccctcct gcccctcctg cccctcctcc
     6541 tgctcctgcc cctcctgctc ctgcccctcc tgcccctcct gcccctcctg ctcctgcccc
     6601 tcctcctgct cctgcccctc ctgcccctcc tgcccctcct cctgctcctg cccctcctgc
     6661 ccctcctcct gctcctgccc ctcctcctgc tcctgcccct cctgcccctc ctgcccctcc
     6721 tcctgctcct gcccctcctg cccctcctcc tgctcctgcc cctcctcctg ctcctgcccc
     6781 tcctgcccct cctgcccctc ctcctgctcc tgcccctcct cctgctcctg cccctcctgc
     6841 ccctcctgcc cctcctgccc ctcctcctgc tcctgcccct cctcctgctc ctgcccctcc
     6901 tgctcctgcc cctcccgctc ctgctcctgc tcctgttcca ccgtgggtcc ctttgcagcc
     6961 aatgcaactt ggacgttttt ggggtctccg gacaccatct ctatgtcttg gccctgatcc
     7021 tgagccgccc ggggctcctg gtcttccgcc tcctcgtcct cgtcctcttc cccgtcctcg
     7081 tccatggtta tcaccccctc ttctttgagg tccactgccg ccggagcctt ctggtccaga
     7141 tgtgtctccc ttctctccta ggccatttcc aggtcctgta cctggcccct cgtcagacat
     7201 gattcacact aaaagagatc aatagacatc tttattagac gacgctcagt gaatacaggg
     7261 agtgcagact cctgccccct ccaacagccc ccccaccctc atccccttca tggtcgctgt
     7321 cagacagatc caggtctgaa aattccccat cctccgaacc atcctcgtcc tcatcaccaa
     7381 ttactcgcag cccggaaaac tcccgctgaa catcctcaag atttgcgtcc tgagcctcaa
     7441 gccaggcctc aaattcctcg tccccctttt tgctggacgg tagggatggg gattctcggg
     7501 acccctcctc ttcctcttca aggtcaccag acagagatgc tactggggca acggaagaaa
     7561 agctgggtgc ggcctgtgag gatcagctta tcgatgataa gctgtcaaac atgagaattc
     7621 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     7681 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     7741 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     7801 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     7861 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     7921 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     7981 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     8041 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     8101 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     8161 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     8221 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     8281 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     8341 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     8401 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     8461 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     8521 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     8581 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     8641 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     8701 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     8761 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8821 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     8881 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     8941 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     9001 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     9061 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     9121 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     9181 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     9241 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     9301 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     9361 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     9421 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     9481 cttttgctgg ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc
     9541 ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat
     9601 ccagcctcgc gtcgaactag atgatccggc tgtggaatgt gtgtcagtta gggtgtggaa
     9661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     9721 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     9781 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     9841 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     9901 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga gggtgaccgc
     9961 cacgaccggt gccgccacca tcccctgacc cacgcccctg acccctcaca aggagacgac
    10021 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
    10081 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
    10141 ccgaccgcca catcgaacgc gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
    10201 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
    10261 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
    10321 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
    10381 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
    10441 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
    10501 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
    10561 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
    10621 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga cccggtccga
    10681 cgcggcccga cgggtccgag gggggtcgac ctcgaaactt gtttattgca gcttataatg
    10741 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
    10801 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatccgaac
    10861 cccttcctcg accaattctc atgtttgaca gcttatcatc gcagatccgg gcaacgttgt
    10921 tgccattgct cgaccaattc tcatgtttga cagcttatca tcgcagatcc gtatggtgca
    10981 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
    11041 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
    11101 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
    11161 gggccagata tacgcgtatc tgaggggact agggtgtgtt taggcgaaaa gcggggcttc
    11221 ggttgtacgc ggttaggagt cccctcagga tatagtagtt tcgcttttgc atagggaggg
    11281 ggaaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
    11341 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
    11401 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
    11461 attccgcatt gcagagatat tgtatttaag tgcctagctc gatacaataa acgccatttg
    11521 accattcacc acattggtgt gcacctccaa gctggccggg gatcctctag ggcctctgag
    11581 ctattccaga agtagtgaag aggctttttt ggaggcctag gcttttgcaa aaagctccgg
    11641 atcgatcctg agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct
    11701 attgtaaaat tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga
    11761 agatgtccct tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc
    11821 tgttgacaac cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac
    11881 tttagcttgc atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag
    11941 tactttctct aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca
    12001 cagttttaga gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat
    12061 tctggctggc gtggaaatat tcttattggt agaaacaact acatcctggt catcatcctg
    12121 cctttctctt tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag
    12181 tccaaaccgg gcccctctgc taaccatgtt catgccttct tctttttcct acagctcctg
    12241 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgta atacgactca
    12301 ctatagggcg aattgatatg tctagattag ataaaagtaa agtgattaac agcgcattag
    12361 agctgcttaa tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag
    12421 gtgtagagca gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct
    12481 tagccattga gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct
    12541 ggcaagattt tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg
    12601 atggagcaaa agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa
    12661 atcaattagc ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca
    12721 gcgctgtggg gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta
    12781 aagaagaaag ggaaacacct actactgata gtatgccgcc attattacga caagctatcg
    12841 aattatttga tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat
    12901 gcggattaga aaaacaactt aaatgtgaaa gtgggtccgc gtacagcgga tcccgggaat
    12961 tcagatctta ttaaagcaga acttgtttat tgcagcttat aatggttaca aataaagcaa
    13021 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    13081 caaactcatc aatgtatctt atcatgtctg gtcgaatcca tggcgcagaa ctggtaggta
    13141 tggaagatcc gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
    13201 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
    13261 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
    13321 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
    13381 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
    13441 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
    13501 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
    13561 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
    13621 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
    13681 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
    13741 tccctatcag tgatagagat ctccctatca gtgatagaga tcgtcgacga gctcgtttag
    13801 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
    13861 gggaccgatc cagcctccgg actctagcgt ttaaactta
//