LOCUS       sequence-462110-       15578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Halo tagged human LRRK2 with LIR1 and LIR2 point mutations
            in mammalian cells. Stable expression with piggybac transposase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15578)
  AUTHORS   Bentley-DeSousa A, Roczniak-Ferguson A, Ferguson SM
  TITLE     A STING-CASM-GABARAP pathway activates LRRK2 at lysosomes.
  JOURNAL   J Cell Biol. 2025 Feb 3;224(2):e202310150. doi: 
            10.1083/jcb.202310150. Epub 2025 Jan 15.
  PUBMED    39812709
REFERENCE   2  (bases 1 to 15578)
  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"; journalName: "J Cell 
            Biol."; date: "2025-02-3"; volume: "224(2):e202310150. doi"; pages: 
            " 10.1083/jcb.202310150. Epub 2025 Jan 15"
COMMENT     Sequence Label: pPB-EF1A-HALO-hLRRK2 (LIR1+2 mutant WEVL -> AE...
FEATURES             Location/Qualifiers
     source          1..15578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..39
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    316..336
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        337..1515
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          568..1506
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1463..1483
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1515..1539
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1540..1549
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1546..2433
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     protein_bind    complement(10066..10090)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(10168..10187)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    10233..10288
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(10287..10306)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        10643..11022
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11023..11226
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11177..11197
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             11259..11858
                     /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(11259..11278)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     11755..11775
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(11896..11913)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11902..12126
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   12476..12538
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(12604..12623)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(12605..12623)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(12630..12647)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12630..12646)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12639..12661)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      12788..13243
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12875..12894)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13085..13106
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        13269..13373
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(13374..13442,13443..14234)
                     /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(13592..13611)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      14405..14993
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14894..14913
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     15147..15164
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    15281..15302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(15317..15322,15323..15340,15341..15347)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15355..15371
                     /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     15360..15382
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     15379..15395
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15379..15395
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        15416..15434
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc
       61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga
      121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc
      181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg
      241 ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct
      301 tgttatagat atcatcaact ttgtatagaa aagttgggct ccggtgcccg tcagtgggca
      361 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
      421 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
      481 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
      541 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
      601 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg
      661 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa
      721 ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg
      781 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
      841 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
      901 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
      961 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
     1021 gggtagtctc aagctggccg gcctgctctg gtgcctggtc tcgcgccgcc gtgtatcgcc
     1081 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
     1141 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg
     1201 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
     1261 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg
     1321 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
     1381 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
     1441 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
     1501 tttcaggtgt cgtgacaagt ttgtacaaaa aagcaggctg ccaccatgga aatcggtact
     1561 ggctttccat tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat
     1621 gttggtccgc gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac
     1681 gtgtggcgca acatcatccc gcatgttgca ccgacccatc gctgcattgc tccagacctg
     1741 atcggtatgg gcaaatccga caaaccagac ctgggttatt tcttcgacga ccacgtccgc
     1801 ttcatggatg ccttcatcga agccctgggt ctggaagagg tcgtcctggt cattcacgac
     1861 tggggctccg ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt
     1921 gcatttatgg agttcatccg ccctatcccg acctgggacg aatggccaga atttgcccgc
     1981 gagaccttcc aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac
     2041 gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg
     2101 gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca
     2161 aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg
     2221 gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg
     2281 atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac
     2341 atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc
     2401 gcgcgctggc tgtccacgct cgagatttcc ggctctggtg gcggaggctc gggcggaggt
     2461 gggtcgggtg gcggcggatc aatggctagt ggcagctgtc aggggtgcga agaggacgag
     2521 gaaactctga agaagttgat agtcaggctg aacaatgtcc aggaaggaaa acagatagaa
     2581 acgctggtcc aaatcctgga ggatctgctg gtgttcacgt actccgagcg cgcctccaag
     2641 ttatttcaag gcaaaaatat ccatgtgcct ctgttgatcg tcttggactc ctatatgaga
     2701 gtcgcgagtg tgcagcaggt gggttggtca cttctgtgca aattaataga agtctgtcca
     2761 ggtacaatgc aaagcttaat gggaccccag gatgttggaa atgatgcgga agtcgctggt
     2821 gttcaccaat tgattcttaa aatgctaaca gttcataatg ccagtgtaaa cttgtcagtg
     2881 attggactga agaccttaga tctcctccta acttcaggta aaatcacctt gctgatattg
     2941 gatgaagaaa gtgatatttt catgttaatt tttgatgcca tgcactcatt tccagccaat
     3001 gatgaagtcc agaaacttgg atgcaaagct ttacatgtgc tgtttgagag agtctcagag
     3061 gagcaactga ctgaatttgt tgagaacaaa gattatatga tattgttaag tgcgttaaca
     3121 aattttaaag atgaagagga aattgtgctt catgtgctgc attgtttaca ttccctagcg
     3181 attccttgca ataatgtgga agtcctcatg agtggcaatg tcaggtgtta taatattgtg
     3241 gtggaagcta tgaaagcatt ccctatgagt gaaagaattc aagaagtgag ttgctgtttg
     3301 ctccataggc ttacattagg taattttttc aatatcctgg tattaaacga agtccatgag
     3361 tttgtggtga aagctgtgca gcagtaccca gagaatgcag cattgcagat ctcagcgctc
     3421 agctgtttgg ccctcctcac tgagactatt ttcttaaatc aagatttaga ggaaaagaat
     3481 gagaatcaag agaatgatga tgagggggaa gaagataaat tgttttggct ggaagcctgt
     3541 tacaaagcat taacgtggca tagaaagaac aagcacgtgc aggaggccgc atgctgggca
     3601 ctaaataatc tccttatgta ccaaaacagt ttacatgaga agattggaga tgaagatggc
     3661 catttcccag ctcataggga agtgatgctc tccatgctga tgcattcttc atcaaaggaa
     3721 gttttccagg catctgcgaa tgcattgtca actctcttag aacaaaatgt taatttcaga
     3781 aaaatactgt tatcaaaagg aatacacctg aatgttttgg agttaatgca gaagcatata
     3841 cattctcctg aagtggctga aagtggctgt aaaatgctaa atcatctttt tgaaggaagc
     3901 aacacttccc tggatataat ggcagcagtg gtccccaaaa tactaacagt tatgaaacgt
     3961 catgagacat cattaccagt gcagctggag gcgcttcgag ctattttaca ttttatagtg
     4021 cctggcatgc cagaagaatc cagggaggat acagaatttc atcataagct aaatatggtt
     4081 aaaaaacagt gtttcaagaa tgatattcac aaactggtcc tagcagcttt gaacaggttc
     4141 attggaaatc ctgggattca gaaatgtgga ttaaaagtaa tttcttctat tgtacatttt
     4201 cctgatgcat tagagatgtt atccctggaa ggtgctatgg attcagtgct tcacacactg
     4261 cagatgtatc cagatgacca agaaattcag tgtctgggtt taagtcttat aggatacttg
     4321 attacaaaga agaatgtgtt cataggaact ggacatctgc tggcaaaaat tctggtttcc
     4381 agcttatacc gatttaagga tgttgctgaa atacagacta aaggatttca gacaatctta
     4441 gcaatcctca aattgtcagc atctttttct aagctgctgg tgcatcattc atttgactta
     4501 gtaatattcc atcaaatgtc ttccaatatc atggaacaaa aggatcaaca gtttctaaac
     4561 ctctgttgca agtgttttgc aaaagtagct atggatgatt acttaaaaaa tgtgatgcta
     4621 gagagagcgt gtgatcagaa taacagcatc atggttgaat gcttgcttct attgggagca
     4681 gatgccaatc aagcaaagga gggatcttct ttaatttgtc aggtatgtga gaaagagagc
     4741 agtcccaaat tggtggaact cttactgaat agtggatctc gtgaacaaga tgtacgaaaa
     4801 gcgttgacga taagcattgg gaaaggtgac agccagatca tcagcttgct cttaaggagg
     4861 ctggccctgg atgtggccaa caatagcatt tgccttggag gattttgtat aggaaaagtt
     4921 gaaccttctt ggcttggtcc tttatttcca gataagactt ctaatttaag gaaacaaaca
     4981 aatatagcat ctacactagc aagaatggtg atcagatatc agatgaaaag tgctgtggaa
     5041 gaaggaacag cctcaggcag cgatggaaat ttttctgaag atgtgctgtc taaatttgat
     5101 gaagcgacct ttgctcctga ctcttctatg gacagtgtgt ttgctcaaag tgatgacctg
     5161 gatagtgaag gaagtgaagg ctcatttctt gtgaaaaaga aatctaattc aattagtgta
     5221 ggagaatttt accgagatgc cgtattacag cgttgctcac caaatttgca aagacattcc
     5281 aattccttgg ggcccatttt tgatcatgaa gatttactga agcgaaaaag aaaaatatta
     5341 tcttcagatg attcactcag gtcatcaaaa cttcaatccc atatgaggca ttcagacagc
     5401 atttcttctc tggcttctga gagagaatat attacatcac tagacctttc agcaaatgaa
     5461 ctaagagata ttgatgccct aagccagaaa tgctgtataa gtgttcattt ggagcatctt
     5521 gaaaagctgg agcttcacca gaatgcactc acgagctttc cacaacagct atgtgaaact
     5581 ctgaagagtt tgacacattt ggacttgcac agtaataaat ttacatcatt tccttcttat
     5641 ttgttgaaaa tgagttgtat tgctaatctt gatgtctctc gaaatgacat tggaccctca
     5701 gtggttttag atcctacagt gaaatgtcca actctgaaac agtttaacct gtcatataac
     5761 cagctgtctt ttgtacctga gaacctcact gatgtggtag agaaactgga gcagctcatt
     5821 ttagaaggaa ataaaatatc agggatatgc tcccccttga gactgaagga actgaagatt
     5881 ttaaacctta gtaagaacca catttcatcc ctatcagaga actttcttga ggcttgtcct
     5941 aaagtggaga gtttcagtgc cagaatgaat tttcttgctg ctatgccttt cttgcctcct
     6001 tctatgacaa tcctaaaatt atctcagaac aaattttcct gtattccaga agcaatttta
     6061 aatcttccac acttgcggtc tttagatatg agcagcaatg atattcagta cctaccaggt
     6121 cccgcacact ggaaatcttt gaacttaagg gaactcttat ttagccataa tcagatcagc
     6181 atcttggact tgagtgaaaa agcatattta tggtctagag tagagaaact gcatctttct
     6241 cacaataaac tgaaagagat tcctcctgag attggctgtc ttgaaaatct gacatctctg
     6301 gatgtcagtt acaacttgga actaagatcc tttcccaatg aaatggggaa attaagcaaa
     6361 atatgggatc ttcctttgga tgaactgcat cttaactttg attttaaaca tataggatgt
     6421 aaagccaaag acatcataag gtttcttcaa cagcgattaa aaaaggctgt gccttataac
     6481 cgaatgaaac ttatgattgt gggaaatact gggagtggta aaaccacctt attgcagcaa
     6541 ttaatgaaaa ccaagaaatc agatcttgga atgcaaagtg ccacagttgg catagatgtg
     6601 aaagactggc ctatccaaat aagagacaaa agaaagagag atctcgtcct aaatgtgtgg
     6661 gattttgcag gtcgtgagga attctatagt actcatcccc attttatgac gcagcgagca
     6721 ttgtaccttg ctgtctatga cctcagcaag ggacaggctg aagttgatgc catgaagcct
     6781 tggctcttca atataaaggc tcgcgcttct tcttcccctg tgattctcgt tggcacacat
     6841 ttggatgttt ctgatgagaa gcaacgcaaa gcctgcatga gtaaaatcac caaggaactc
     6901 ctgaataagc gagggttccc tgccatacga gattaccact ttgtgaatgc caccgaggaa
     6961 tctgatgctt tggcaaaact tcggaaaacc atcataaacg agagccttaa tttcaagatc
     7021 cgagatcagc ttgttgttgg acagctgatt ccagactgct atgtagaact tgaaaaaatc
     7081 attttatcgg agcgtaaaaa tgtgccaatt gaatttcccg taattgaccg gaaacgatta
     7141 ttacaactag tgagagaaaa tcagctgcag ttagatgaaa atgagcttcc tcacgcagtt
     7201 cactttctaa atgaatcagg agtccttctt cattttcaag acccagcact gcagttaagt
     7261 gacttgtact ttgtggaacc caagtggctt tgtaaaatca tggcacagat tttgacagtg
     7321 aaagtggaag gttgtccaaa acaccctaag ggcattattt cgcgtagaga tgtggaaaaa
     7381 tttctttcaa aaaaaaggaa atttccaaag aactacatgt cacagtattt taagctccta
     7441 gaaaaattcc agattgcttt gccaatagga gaagaatatt tgctggttcc aagcagtttg
     7501 tctgaccaca ggcctgtgat agagcttccc cattgtgaga actctgaaat tatcatccga
     7561 ctatatgaaa tgccttattt tccaatggga ttttggtcaa gattaatcaa tcgattactt
     7621 gagatttcac cttacatgct ttcagggaga gaacgagcac ttcgcccaaa cagaatgtat
     7681 tggcgacaag gcatttactt aaattggtct cctgaagctt attgtctggt aggatctgaa
     7741 gtcttagaca atcatccaga gagtttctta aaaattacag ttccttcttg tagaaaaggc
     7801 tgtattcttt tgggccaagt tgtggaccac attgattctc tcatggaaga atggtttcct
     7861 gggttgctgg agattgatat ttgtggtgaa ggagaaactc tgttgaagaa atgggcatta
     7921 tatagtttta atgatggtga agaacatcaa aaaatcttac ttgatgactt gatgaagaaa
     7981 gcagaggaag gagatctctt agtaaatcca gatcaaccaa ggctcaccat tccaatatct
     8041 cagattgccc ctgacttgat tttggctgac ctgcctagaa atattatgtt gaataatgat
     8101 gagttggaat ttgaacaagc tccagagttt ctcctaggtg atggcagttt tggatcagtt
     8161 taccgagcag cctatgaagg agaagaagtg gctgtgaaga tttttaataa acatacatca
     8221 ctcaggctgt taagacaaga gcttgtggtg ctttgccacc tccaccaccc cagtttgata
     8281 tctttgctgg cagctgggat tcgtccccgg atgttggtga tggagttagc ctccaagggt
     8341 tccttggatc gcctgcttca gcaggacaaa gccagcctca ctagaaccct acagcacagg
     8401 attgcactcc acgtagctga tggtttgaga tacctccact cagccatgat tatataccga
     8461 gacctgaaac cccacaatgt gctgcttttc acactgtatc ccaatgctgc catcattgca
     8521 aagattgctg actacggcat tgctcagtac tgctgtagaa tggggataaa aacatcagag
     8581 ggcacaccag ggtttcgtgc acctgaagtt gccagaggaa atgtcattta taaccaacag
     8641 gctgatgttt attcatttgg tttactactc tatgacattt tgacaactgg aggtagaata
     8701 gtagagggtt tgaagtttcc aaatgagttt gatgaattag aaatacaagg aaaattacct
     8761 gatccagtta aagaatatgg ttgtgcccca tggcctatgg ttgagaaatt aattaaacag
     8821 tgtttgaaag aaaatcctca agaaaggcct acttctgccc aggtctttga cattttgaat
     8881 tcagctgaat tagtctgtct gacgagacgc attttattac ctaaaaacgt aattgttgaa
     8941 tgcatggttg ctacacatca caacagcagg aatgcaagca tttggctggg ctgtgggcac
     9001 accgacagag gacagctctc atttcttgac ttaaatactg aaggatacac ttctgaggaa
     9061 gttgctgata gtagaatatt gtgcttagcc ttggtgcatc ttcctgttga aaaggaaagc
     9121 tggattgtgt ctgggacaca gtctggtact ctcctggtca tcaataccga agatgggaaa
     9181 aagagacata ccctagaaaa gatgactgat tctgtcactt gtttgtattg caattccttt
     9241 tccaagcaaa gcaaacaaaa aaattttctt ttggttggaa ccgctgatgg caagttagca
     9301 atttttgaag ataagactgt taagcttaaa ggagctgctc ctttgaagat actaaatata
     9361 ggaaatgtca gtactccatt gatgtgtttg agtgaatcca caaattcaac ggaaagaaat
     9421 gtaatgtggg gaggatgtgg cacaaagatt ttctcctttt ctaatgattt caccattcag
     9481 aaactcattg agacaagaac aagccaactg ttttcttatg cagctttcag tgattccaac
     9541 atcataacag tggtggtaga cactgctctc tatattgcta agcaaaatag ccctgttgtg
     9601 gaagtgtggg ataagaaaac tgaaaaactc tgtggactaa tagactgcgt gcacttttta
     9661 agggaggtaa tggtaaaaga aaacaaggaa tcaaaacaca aaatgtctta ttctgggaga
     9721 gtgaaaaccc tctgccttca gaagaacact gctctttgga taggaactgg aggaggccat
     9781 attttactcc tggatctttc aactcgtcga cttatacgtg taatttacaa cttttgtaat
     9841 tcggtcagag tcatgatgac agcacagcta ggaagcctta aaaatgtcat gctggtattg
     9901 ggctacaacc ggaaaaatac tgaaggtaca caaaagcaga aagagataca atcttgcttg
     9961 accgtttggg acatcaatct tccacatgaa gtgcaaaatt tagaaaaaca cattgaagtg
    10021 agaaaagaat tagctgaaaa aatgagacga acatctgttg agtaaaccca gctttcttgt
    10081 acaaagtggt gatcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
    10141 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
    10201 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
    10261 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
    10321 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
    10381 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
    10441 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
    10501 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
    10561 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga
    10621 cctgcagccc aagcttcgcg ttgacattga ttattgacta gttattaata gtaatcaatt
    10681 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
    10741 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
    10801 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
    10861 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
    10921 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
    10981 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
    11041 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
    11101 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
    11161 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
    11221 agagctctct ggctaactag agaacccact gcgccaccat gaccgagtac aagcccacgg
    11281 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
    11341 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
    11401 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
    11461 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
    11521 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
    11581 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
    11641 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
    11701 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
    11761 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
    11821 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgact cgagtctaga gggcccgttt
    11881 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
    11941 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
    12001 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
    12061 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
    12121 ctatggctcg agttaattaa cgagagcata atattgatat gtgccaaagt tgtttctgac
    12181 tgactaataa gtataatttg tttctattat gtataggtta agctaattac ttattttata
    12241 atacaacatg actgttttta aagtacaaaa taagtttatt tttgtaaaag agagaatgtt
    12301 taaaagtttt gttactttat agaagaaatt ttgagttttt gttttttttt aataaataaa
    12361 taaacataaa taaattgttt gttgaattta ttattagtat gtaagtgtaa atataataaa
    12421 acttaatatc tattcaaatt aataaataaa cctcgatata cagaccgata aaacacatgc
    12481 gtcaatttta cgcatgatta tctttaacgt acgtcacaat atgattatct ttctagggtt
    12541 aaataatagt ttctaatttt tttattattc agcctgctgt cgtgaatacc gagctccaat
    12601 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
    12661 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
    12721 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
    12781 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
    12841 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
    12901 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
    12961 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
    13021 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
    13081 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
    13141 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
    13201 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
    13261 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
    13321 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
    13381 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
    13441 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
    13501 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
    13561 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
    13621 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
    13681 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
    13741 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
    13801 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
    13861 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
    13921 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
    13981 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
    14041 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
    14101 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
    14161 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
    14221 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
    14281 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
    14341 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
    14401 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
    14461 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
    14521 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
    14581 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
    14641 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
    14701 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
    14761 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
    14821 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
    14881 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
    14941 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
    15001 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
    15061 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
    15121 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
    15181 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
    15241 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
    15301 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
    15361 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctcgaaatta
    15421 accctcacta aagggaacaa aagctggtac ctcgcgcgac ttggtttgcc attctttagc
    15481 gcgcgtcgcg tcacacagct tggccacaat gtggtttttg tcaaacgaag attctatgac
    15541 gtgtttaaag tttaggtcga gtaaagcgca aatctttt
//
