LOCUS       sequence-439114-       10010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-tagged human TBK1 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-EGFP
FEATURES             Location/Qualifiers
     source          1..10010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..229
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     180..200
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             262..861
                     /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(262..281)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     758..778
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(899..916)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    905..1129
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1479..1541
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1607..1626)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1608..1626)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1633..1650)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1633..1649)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1642..1664)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1791..2246
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1878..1897)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2088..2109
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2272..2376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2377..2445,2446..3237)
                     /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     complement(2595..2614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3408..3996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3897..3916
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4150..4167
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4284..4305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4320..4325,4326..4343,4344..4350)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4358..4374
                     /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)."
     primer_bind     4363..4385
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4382..4398
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4382..4398
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4419..4437
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   4586..4620
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4897..4917
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        4918..6096
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5149..6087
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6044..6064
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    6096..6120
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(8362..8364,8365..9078)
                     /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             join(8362..8364,8365..9075)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8404..8425)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8665..8684)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9012..9033
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(9079..9103)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(9181..9200)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    9246..9301
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9300..9319)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        join(9656..10010,1..25)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
       61 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      121 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      181 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      241 tagagaaccc actgcgccac catgaccgag tacaagccca cggtgcgcct cgccacccgc
      301 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
      361 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
      421 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
      481 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
      541 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
      601 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
      661 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
      721 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
      781 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
      841 acccgcaagc ccggtgcctg actcgagtct agagggcccg tttaaacccg ctgatcagcc
      901 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
      961 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     1021 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     1081 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc tcgagttaat
     1141 taacgagagc ataatattga tatgtgccaa agttgtttct gactgactaa taagtataat
     1201 ttgtttctat tatgtatagg ttaagctaat tacttatttt ataatacaac atgactgttt
     1261 ttaaagtaca aaataagttt atttttgtaa aagagagaat gtttaaaagt tttgttactt
     1321 tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat aaataaattg
     1381 tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat atctattcaa
     1441 attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt ttacgcatga
     1501 ttatctttaa cgtacgtcac aatatgatta tctttctagg gttaaataat agtttctaat
     1561 ttttttatta ttcagcctgc tgtcgtgaat accgagctcc aattcgccct atagtgagtc
     1621 gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     1681 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     1741 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     1801 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1861 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1921 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1981 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2041 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2101 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2161 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2221 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     2281 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2341 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2401 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2461 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2521 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2581 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2641 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2701 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2761 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2821 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2881 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2941 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3001 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3061 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3121 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3181 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3241 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3301 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3361 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3421 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3481 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3541 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3601 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3661 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3721 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3781 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3841 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3901 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3961 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4021 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4081 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4141 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4201 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4261 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     4321 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     4381 acaggaaaca gctatgacca tgattacgcc aagctcgaaa ttaaccctca ctaaagggaa
     4441 caaaagctgg tacctcgcgc gacttggttt gccattcttt agcgcgcgtc gcgtcacaca
     4501 gcttggccac aatgtggttt ttgtcaaacg aagattctat gacgtgttta aagtttaggt
     4561 cgagtaaagc gcaaatcttt tttaacccta gaaagatagt ctgcgtaaaa ttgacgcatg
     4621 cattcttgaa atattgctct ctctttctaa atagcgcgaa tccgtcgctg tgcatttagg
     4681 acatctcagt cgccgcttgg agctcccgtg aggcgtgctt gtcaatgcgg taagtgtcac
     4741 tgattttgaa ctataacgac cgcgtgagtc aaaatgacgc atgattatct tttacgtgac
     4801 ttttaagatt taactcatac gataattata ttgttatttc atgttctact tacgtgataa
     4861 cttattatat atatattttc ttgttataga tatcatcaac tttgtataga aaagttgggc
     4921 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     4981 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     5041 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     5101 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
     5161 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
     5221 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
     5281 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
     5341 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
     5401 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
     5461 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
     5521 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
     5581 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggt
     5641 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
     5701 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     5761 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     5821 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     5881 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     5941 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     6001 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     6061 gttcaaagtt tttttcttcc atttcaggtg tcgtgacaag tttgtacaaa aaagcaggct
     6121 gccaccatgc agagcacttc taatcatctg tggcttttat ctgatatttt aggccaagga
     6181 gctactgcaa atgtctttcg tggaagacat aagaaaactg gtgatttatt tgctatcaaa
     6241 gtatttaata acataagctt ccttcgtcca gtggatgttc aaatgagaga atttgaagtg
     6301 ttgaaaaaac tcaatcacaa aaatattgtc aaattatttg ctattgaaga ggagacaaca
     6361 acaagacata aagtacttat tatggaattt tgtccatgtg ggagtttata cactgtttta
     6421 gaagaacctt ctaatgccta tggactacca gaatctgaat tcttaattgt tttgcgagat
     6481 gtggtgggtg gaatgaatca tctacgagag aatggtatag tgcaccgtga tatcaagcca
     6541 ggaaatatca tgcgtgttat aggggaagat ggacagtctg tgtacaaact cacagatttt
     6601 ggtgcagcta gagaattaga agatgatgag cagtttgttt ctctgtatgg cacagaagaa
     6661 tatttgcacc ctgatatgta tgagagagca gtgctaagaa aagatcatca gaagaaatat
     6721 ggagcaacag ttgatctttg gagcattggg gtaacatttt accatgcagc tactggatca
     6781 ctgccattta gaccctttga agggcctcgt aggaataaag aagtgatgta taaaataatt
     6841 acaggaaagc cttctggtgc aatatctgga gtacagaaag cagaaaatgg accaattgac
     6901 tggagtggag acatgcctgt ttcttgcagt ctttctcggg gtcttcaggt tctacttacc
     6961 cctgttcttg caaacatcct tgaagcagat caggaaaagt gttggggttt tgaccagttt
     7021 tttgcagaaa ctagtgatat acttcaccga atggtaattc atgttttttc gctacaacaa
     7081 atgacagctc ataagattta tattcatagc tataatactg ctactatatt tcatgaactg
     7141 gtatataaac aaaccaaaat tatttcttca aatcaagaac ttatctacga agggcgacgc
     7201 ttagtcttag aacctggaag gctggcacaa catttcccta aaactactga ggaaaaccct
     7261 atatttgtag taagccggga acctctgaat accataggat taatatatga aaaaatttcc
     7321 ctccctaaag tacatccacg ttatgattta gacggggatg ctagcatggc taaggcaata
     7381 acaggggttg tgtgttatgc ctgcagaatt gccagtacct tactgcttta tcaggaatta
     7441 atgcgaaagg ggatacgatg gctgattgaa ttaattaaag atgattacaa tgaaactgtt
     7501 cacaaaaaga cagaagttgt gatcacattg gatttctgta tcagaaacat tgaaaaaact
     7561 gtgaaagtat atgaaaagtt gatgaagatc aacctggaag cggcagagtt aggtgaaatt
     7621 tcagacatac acaccaaatt gttgagactt tccagttctc agggaacaat agaaaccagt
     7681 cttcaggata tcgacagcag attatctcca ggtggatcac tggcagacgc atgggcacat
     7741 caagaaggca ctcatccgaa agacagaaat gtagaaaaac tacaagtcct gttaaattgc
     7801 atgacagaga tttactatca gttcaaaaaa gacaaagcag aacgtagatt agcttataat
     7861 gaagaacaaa tccacaaatt tgataagcaa aaactgtatt accatgccac aaaagctatg
     7921 acgcacttta cagatgaatg tgttaaaaag tatgaggcat ttttgaataa gtcagaagaa
     7981 tggataagaa agatgcttca tcttaggaaa cagttattat cgctgactaa tcagtgtttt
     8041 gatattgaag aagaagtatc aaaatatcaa gaatatacta atgagttaca agaaactctg
     8101 cctcagaaaa tgtttacagc ttccagtgga atcaaacata ccatgacccc aatttatcca
     8161 agttctaaca cattagtaga aatgactctt ggtatgaaga aattaaagga agagatggaa
     8221 ggggtggtta aagaacttgc tgaaaataac cacattttag aaaggtttgg ctctttaacc
     8281 atggatggtg gccttcgcaa cgttgactgt ctttctggtg gcggaggctc gggcggaggt
     8341 gggtcgggtg gcggcggatc agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     8401 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     8461 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     8521 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     8581 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     8641 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     8701 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     8761 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     8821 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     8881 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     8941 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     9001 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     9061 gacgagctgt acaagtagac ccagctttct tgtacaaagt ggtgatcctc aggtgcaggc
     9121 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     9181 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     9241 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     9301 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     9361 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     9421 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     9481 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     9541 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     9601 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttc gcgttgacat
     9661 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     9721 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     9781 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     9841 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     9901 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     9961 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
//
