LOCUS       sequence-439111-       10010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-tagged human TBK1 with E696K mutation in mammalian 
            cells. Compatible with piggybac transposase for genome integration..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10010)
  AUTHORS   Talaia G, Bentley-DeSousa A, Ferguson SM
  TITLE     Lysosomal TBK1 responds to amino acid availability to relieve 
            Rab7-dependent mTORC1 inhibition.
  JOURNAL   EMBO J. 2024 Sep;43(18):3948-3967. doi: 10.1038/s44318-024-00180-8. 
            Epub 2024 Aug 5.
  PUBMED    39103493
REFERENCE   2  (bases 1 to 10010)
  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: 
            "10.1038/s44318-024-00180-8"; journalName: "EMBO J"; date: 
            "2024-09"; volume: "43"; issue: "18"; pages: "3948-3967"
COMMENT     Sequence Label: pPB-EF1A-Puro-hTBK1-E696K-EGFP
FEATURES             Location/Qualifiers
     source          1..10010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1132..1156)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(1156..2334)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1165..2103
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(1188..1208)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(2335..2355)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     repeat_region   2632..2666
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(2815..2833)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2854..2870)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2854..2870)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2867..2889)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2878..2894
                     /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(2902..2908,2909..2926,2927..2932))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2947..2968
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3085..3102)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3256..3844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3336..3355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4015..4806,4807..4875))
                     /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     4638..4657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4876..4980)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5006..5461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5143..5164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5355..5374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5588..5610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5602..5619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5603..5619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5626..5645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5626..5644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   5711..5773
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    6123..6347
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6336..6353
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(6391..6990)
                     /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(6474..6494)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6971..6990
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(7023..7226)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7052..7072)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7227..7606
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7943..7962
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    7961..8016
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     8062..8081
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     protein_bind    8159..8183
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(8184..8897,8898..8900))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(8187..8897,8898..8900))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8229..8250)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     8578..8597
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8837..8858
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 ctcagtagtt ttagggaaat gttgtgccag ccttccaggt tctaagacta agcgtcgccc
       61 ttcgtagata agttcttgat ttgaagaaat aattttggtt tgtttatata ccagttcatg
      121 aaatatagta gcagtattat agctatgaat ataaatctta tgagctgtca tttgttgtag
      181 cgaaaaaaca tgaattacca ttcggtgaag tatatcacta gtttctgcaa aaaactggtc
      241 aaaaccccaa cacttttcct gatctgcttc aaggatgttt gcaagaacag gggtaagtag
      301 aacctgaaga ccccgagaaa gactgcaaga aacaggcatg tctccactcc agtcaattgg
      361 tccattttct gctttctgta ctccagatat tgcaccagaa ggctttcctg taattatttt
      421 atacatcact tctttattcc tacgaggccc ttcaaagggt ctaaatggca gtgatccagt
      481 agctgcatgg taaaatgtta ccccaatgct ccaaagatca actgttgctc catatttctt
      541 ctgatgatct tttcttagca ctgctctctc atacatatca gggtgcaaat attcttctgt
      601 gccatacaga gaaacaaact gctcatcatc ttctaattct ctagctgcac caaaatctgt
      661 gagtttgtac acagactgtc catcttcccc tataacacgc atgatatttc ctggcttgat
      721 atcacggtgc actataccat tctctcgtag atgattcatt ccacccacca catctcgcaa
      781 aacaattaag aattcagatt ctggtagtcc ataggcatta gaaggttctt ctaaaacagt
      841 gtataaactc ccacatggac aaaattccat aataagtact ttatgtcttg ttgttgtctc
      901 ctcttcaata gcaaataatt tgacaatatt tttgtgattg agttttttca acacttcaaa
      961 ttctctcatt tgaacatcca ctggacgaag gaagcttatg ttattaaata ctttgatagc
     1021 aaataaatca ccagttttct tatgtcttcc acgaaagaca tttgcagtag ctccttggcc
     1081 taaaatatca gataaaagcc acagatgatt agaagtgctc tgcatggtgg cagcctgctt
     1141 ttttgtacaa acttgtcacg acacctgaaa tggaagaaaa aaactttgaa ccactgtctg
     1201 aggcttgaga atgaaccaag atccaaactc aaaaagggca aattccaagg agaattacat
     1261 caagtgccaa gctggcctaa cttcagtctc cacccactca gtgtggggaa actccatcgc
     1321 ataaaacccc tccccccaac ctaaagacga cgtactccaa aagctcgaga actaatcgag
     1381 gtgcctggac ggcgcccggt actccgtgga gtcacatgaa gcgacggctg aggacggaaa
     1441 ggcccttttc ctttgtgtgg gtgactcacc cgcccgctct cccgagcgcc gcgtcctcca
     1501 ttttgagctc cctgcagcag ggccgggaag cggccatctt tccgctcacg caactggtgc
     1561 cgaccgggcc agccttgccg cccagggcgg ggcgatacac ggcggcgcga gaccaggcac
     1621 cagagcaggc cggccagctt gagactaccc ccgtccgatt ctcggtggcc gcgctcgcag
     1681 gccccgcctc gccgaacatg tgcgctggga cgcacgggcc ccgtcgccgc ccgcggcccc
     1741 aaaaaccgaa ataccagtgt gcagatcttg gcccgcattt acaagactat cttgccagaa
     1801 aaaaagcgtc gcagcaggtc atcaaaaatt ttaaatggct agagacttat cgaaagcagc
     1861 gagacaggcg cgaaggtgcc accagattcg cacgcggcgg ccccagcgcc caggccaggc
     1921 ctcaactcaa gcacgaggcg aaggggctcc ttaagcgcaa ggcctcgaac tctcccaccc
     1981 acttccaacc cgaagctcgg gatcaagaat cacgtactgc agccaggtgg aagtaattca
     2041 aggcacgcaa gggccataac ccgtaaagag gccaggcccg cgggaaccac acacggcact
     2101 tacctgtgtt ctggcggcaa acccgttgcg aaaaagaacg ttcacggcga ctactgcact
     2161 tatatacggt tctcccccac cctcgggaaa aaggcggagc cagtacacga catcactttc
     2221 ccagtttacc ccgcgccacc ttctctaggc accggttcaa ttgccgaccc ctccccccaa
     2281 cttctcgggg actgtgggcg atgtgcgctc tgcccactga cgggcaccgg agcccaactt
     2341 ttctatacaa agttgatgat atctataaca agaaaatata tatataataa gttatcacgt
     2401 aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa
     2461 aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt
     2521 accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc
     2581 acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat gcatgcgtca
     2641 attttacgca gactatcttt ctagggttaa aaaagatttg cgctttactc gacctaaact
     2701 ttaaacacgt catagaatct tcgtttgaca aaaaccacat tgtggccaag ctgtgtgacg
     2761 cgacgcgcgc taaagaatgg caaaccaagt cgcgcgaggt accagctttt gttcccttta
     2821 gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2881 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2941 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3001 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3061 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3661 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4981 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     5041 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5101 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5161 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5221 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5281 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5341 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5401 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5461 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     5521 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     5581 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     5641 tagggcgaat tggagctcgg tattcacgac agcaggctga ataataaaaa aattagaaac
     5701 tattatttaa ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgcgta
     5761 aaattgacgc atgtgtttta tcggtctgta tatcgaggtt tatttattaa tttgaataga
     5821 tattaagttt tattatattt acacttacat actaataata aattcaacaa acaatttatt
     5881 tatgtttatt tatttattaa aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa
     5941 aacttttaaa cattctctct tttacaaaaa taaacttatt ttgtacttta aaaacagtca
     6001 tgttgtatta taaaataagt aattagctta acctatacat aatagaaaca aattatactt
     6061 attagtcagt cagaaacaac tttggcacat atcaatatta tgctctcgtt aattaactcg
     6121 agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat cctccccctt
     6181 gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc
     6241 aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg
     6301 cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aggctgatca
     6361 gcgggtttaa acgggccctc tagactcgag tcaggcaccg ggcttgcggg tcatgcacca
     6421 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
     6481 gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc
     6541 cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
     6601 gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
     6661 ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
     6721 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc
     6781 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc
     6841 ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg
     6901 gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc
     6961 gaggcgcacc gtgggcttgt actcggtcat ggtggcgcag tgggttctct agttagccag
     7021 agagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     7081 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     7141 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     7201 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     7261 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     7321 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     7381 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     7441 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     7501 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     7561 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcaacg cgaagcttgg
     7621 gctgcaggtc gagggatctc cataagagaa gagggacagc tatgactggg agtagtcagg
     7681 agaggaggaa aaatctggct agtaaaacat gtaaggaaaa ttttagggat gttaaagaaa
     7741 aaaataacac aaaacaaaat ataaaaaaaa tctaacctca agtcaaggct tttctatgga
     7801 ataaggaatg gacagcaggg ggctgtttca tatactgatg acctctttat agccaacctt
     7861 tgttcatggc agccagcata tgggcatatg ttgccaaact ctaaaccaaa tactcattct
     7921 gatgttttaa atgatttgcc ctcccatatg tccttccgag tgagagacac aaaaaattcc
     7981 aacacactat tgcaatgaaa ataaatttcc tttattagcc agaagtcaga tgctcaaggg
     8041 gcttcatgat gtccccataa tttttggcag agggaaaaag atctcagtgg tatttgtgag
     8101 ccagggcatt ggccacacca gccaccacct tctgataggc agcctgcacc tgaggatcac
     8161 cactttgtac aagaaagctg ggtctacttg tacagctcgt ccatgccgag agtgatcccg
     8221 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     8281 agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     8341 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
     8401 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
     8461 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     8521 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     8581 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     8641 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     8701 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     8761 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     8821 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     8881 agctcctcgc ccttgctcac tgatccgccg ccacccgacc cacctccgcc cgagcctccg
     8941 ccaccagaaa gacagtcaac gttgcgaagg ccaccatcca tggttaaaga gccaaacctt
     9001 tctaaaatgt ggttattttc agcaagttct ttaaccaccc cttccatctt ttcctttaat
     9061 ttcttcatac caagagtcat ttctactaat gtgttagaac ttggataaat tggggtcatg
     9121 gtatgtttga ttccactgga agctgtaaac attttctgag gcagagtttc ttgtaactca
     9181 ttagtatatt cttgatattt tgatacttct tcttcaatat caaaacactg attagtcagc
     9241 gataataact gtttcctaag atgaagcatc tttcttatcc attcttctga cttattcaaa
     9301 aatgcctcat actttttaac acattcatct gtaaagtgcg tcatagcttt tgtggcatgg
     9361 taatacagtt tttgcttatc aaatttgtgg atttgttctt cattataagc taatctacgt
     9421 tctgctttgt cttttttgaa ctgatagtaa atctctgtca tgcaatttaa caggacttgt
     9481 agtttttcta catttctgtc tttcggatga gtgccttctt gatgtgccca tgcgtctgcc
     9541 agtgatccac ctggagataa tctgctgtcg atatcctgaa gactggtttc tattgttccc
     9601 tgagaactgg aaagtctcaa caatttggtg tgtatgtctg aaatttcacc taactctgcc
     9661 gcttccaggt tgatcttcat caacttttca tatactttca cagttttttc aatgtttctg
     9721 atacagaaat ccaatgtgat cacaacttct gtctttttgt gaacagtttc attgtaatca
     9781 tctttaatta attcaatcag ccatcgtatc ccctttcgca ttaattcctg ataaagcagt
     9841 aaggtactgg caattctgca ggcataacac acaacccctg ttattgcctt agccatgcta
     9901 gcatccccgt ctaaatcata acgtggatgt actttaggga gggaaatttt ttcatatatt
     9961 aatcctatgg tattcagagg ttcccggctt actacaaata tagggttttc
//
