LOCUS       sequence-358081-       12576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac Vector expressing GFP-RFP-LGALS3 R186S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12576)
  AUTHORS   Eapen VV, Swarup S, Hoyer MJ, Paulo JA, Harper JW
  TITLE     Quantitative proteomics reveals the selectivity of ubiquitin-binding
            autophagy receptors in the turnover of damaged lysosomes by 
            lysophagy.
  JOURNAL   eLife. 2021 Sep 29;10. pii: 72328. doi: 10.7554/eLife.72328.
  PUBMED    34585663
REFERENCE   2  (bases 1 to 12576)
  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: "eLife."; 
            date: "2021-09-29"; volume: "10. pii: 72328. doi"; pages: " 
            10.7554/eLife.72328"
COMMENT     Sequence Label: pAC150-RFP-EGFP-LGALS3 R186S
FEATURES             Location/Qualifiers
     source          1..12576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        162..1340
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          393..1331
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1288..1308
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1401..1425
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1425..2099
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1425..2099
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1555..1573)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1823..1842
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2017..2040
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     regulatory      2112..2121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2118..2120,2121..2123,2124..2834)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2118..2120,2121..2123,2124..2834)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2163..2184)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2424..2443)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(3598..3622)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3675..3716
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3754..3771)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3760..3984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4523..4585
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4623..4642)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4624..4642)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4649..4666)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4649..4665)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4658..4680)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4803..5105
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4984..5003
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5454..6248
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5454..6248
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5508..5527)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6118..6137
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(6498..7289,7290..7358))
                     /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     7121..7140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7482..8070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7971..7990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8224..8241
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8358..8379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8394..8399,8400..8417,8418..8424)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8432..8448
                     /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     8437..8459
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8456..8472
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8456..8472
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8493..8511
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   8550..8584
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    8918..8951
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        9101..9480
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9481..9684
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9635..9655
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9681..9705
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             10869..11864
                     /codon_start=1
                     /product="truncated herpes simplex virus thymidine kinase 
                     (Salomon et al., 1995)"
                     /label=Delta-TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRV
                     LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS
                     SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL
                     PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAV
                     PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFI
                     LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    11900..12034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11937..11956)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11991..12010
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    12063..12096
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
       61 agcctgcaga cacctggggg gatacgggga aaaagcttta ggctgaaaga gagatttaga
      121 atgacagaac tcgatttcat tgcagactgg cgcgcccgtg aggctccggt gcccgtcagt
      181 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
      241 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
      301 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
      361 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
      421 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
      481 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
      541 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
      601 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
      661 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
      721 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
      781 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
      841 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
      901 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
      961 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
     1021 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     1081 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
     1141 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     1201 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     1261 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     1321 ttccatttca ggtgtcgtga agaagcttgg taccgagctc ggatccacta gtccagtgtg
     1381 gtggaattct gcagatatca acaagtttgt acaaaaaagt tggcatggcc tcctccgagg
     1441 acgtcatcaa ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg aacggccacg
     1501 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     1561 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc cctcagttcc
     1621 agtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     1681 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     1741 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     1801 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     1861 ccaccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag atgaggctga
     1921 agctgaagga cggcggccac tacgacgccg aggtcaagac cacctacatg gccaagaagc
     1981 ccgtgcagct gcccggcgcc tacaagaccg acatcaagct ggacatcacc tcccacaacg
     2041 aggactacac catcgtggaa cagtacgagc gcgccgaggg ccgccactcc accggcgccg
     2101 gtggaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2161 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2221 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2281 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     2341 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2401 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2461 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2521 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     2581 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     2641 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2701 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     2761 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2821 acgagctgta caagtccgga ctcagatcta tggcagacaa tttttcgctc catgatgcgt
     2881 tatctgggtc tggaaaccca aaccctcaag gatggcctgg cgcatggggg aaccagcctg
     2941 ctggggcagg gggctaccca ggggcttcct atcctggggc ctaccccggg caggcacccc
     3001 caggggctta tcctggacag gcacctccag gcgcctacca tggagcacct ggagcttatc
     3061 ccggagcacc tgcacctgga gtctacccag ggccacccag cggccctggg gcctacccat
     3121 cttctggaca gccaagtgcc cccggagcct accctgccac tggcccctat ggcgcccctg
     3181 ctgggccact gattgtgcct tataacctgc ctttgcctgg gggagtggtg cctcgcatgc
     3241 tgataacaat tctgggcacg gtgaagccca atgcaaacag aattgcttta gatttccaaa
     3301 gagggaatga tgttgccttc cactttaacc cacgcttcaa tgagaacaac aggagagtca
     3361 ttgtttgcaa tacaaagctg gataataact ggggaaggga agaaagtcag tcggttttcc
     3421 catttgaaag tgggaaacca ttcaaaatac aagtactggt tgaacctgac cacttcaagg
     3481 ttgcagtgaa tgatgctcac ttgttgcagt acaatcatcg ggttaaaaaa ctcaatgaaa
     3541 tcagcaaact gggaatttct ggtgacatag acctcaccag tgcttcatat accatgatac
     3601 caactttctt gtacaaagtg gttgatatcc agcacagtgg cggccgctcg agtctagagg
     3661 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     3721 ccggttaggg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3781 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3841 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3901 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3961 ctggggatgc ggtgggctct atggcggcgc gcctggatcc ccgggtaccg agttgggagc
     4021 tcacggggac agcccccccc caaagccccc agggatgtaa ttacgtccct cccccgctag
     4081 ggggcagcag cgagccgccc ggggctccgc tccggtccgg cgctcccccc gcatccccga
     4141 gccggcagcg tgcggggaca gcccgggcac ggggaaggtg gcacgggatc gctttcctct
     4201 gaacgcttct cgctgctctt tgagcctgca gacacctggg gggatacggg gaaaaagctt
     4261 taggctgaaa gagagattta gaatgacaga actcgatttc attgcagact ggccggccac
     4321 tagtacgcgc cggctcgaca tactagttaa aagttttgtt actttataga agaaattttg
     4381 agtttttgtt tttttttaat aaataaataa acataaataa attgtttgtt gaatttatta
     4441 ttagtatgta agtgtaaata taataaaact taatatctat tcaaattaat aaataaacct
     4501 cgatatacag accgataaaa cacatgcgtc aattttacgc atgattatct ttaacgtacg
     4561 tcacaatatg attatctttc tagggttaaa agggcgaatt cgcggccgct aaattcaatt
     4621 cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg
     4681 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4741 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctatacgtac
     4801 ggcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     4861 agagtgatat tattgacacg ccggggcgac ggatggtgat ccccctggcc agtgcacgtc
     4921 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     4981 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     5041 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     5101 tataaatgtc aggcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga
     5161 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
     5221 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
     5281 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
     5341 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     5401 gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg
     5461 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5521 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5581 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     5641 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5701 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5761 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5821 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5881 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5941 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     6001 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6061 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6121 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6181 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6241 tcttctgaat tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     6301 gcggtatttc acaccgcatc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6361 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagattatc aaaaaggatc
     6421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7381 tgaagcattt atcagggtta ttgtctcatg accaaaatcc cttaacgtga gttttcgttc
     7441 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7501 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7561 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7621 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7681 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7741 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7801 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7861 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7921 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7981 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     8041 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     8101 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8161 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8221 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     8281 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     8341 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     8401 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     8461 aacagctatg accatgatta cgccaagctc agaattaacc ctcactaaag ggactagtcc
     8521 tgcaggttta aacgaattcg cccttttaac cctagaaaga tagtctgcgt aaaattgacg
     8581 catgcattct tgaaatattg ctctctcttt ctaaatagcg cgaatccgtc gctgtgcatt
     8641 taggacatct cagtcgccgc ttggagctcc cgtgaggcgt gcttgtcaat gcggtaagtg
     8701 tcactgattt tgaactataa cgaccgcgtg agtcaaaatg acgcatgatt atcttttacg
     8761 tgacttttaa gatttaactc atacgataat tatattgtta tttcatgttc tacttacgtg
     8821 ataacttatt atatatatat tttcttgtta tagatatcaa ctagaatgct agacctttcg
     8881 tcttcaagaa ttccgatcat attcaataac ccttaatata acttcgtata atgtatgcta
     8941 tacgaagtta ttaggtctga agaggagttt acgtccagcc aagcttagga tcccgggtac
     9001 cgatggccgg gcctcgagga gcttggccca ttgcatacgt tgtatccata tcataatatg
     9061 tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt
     9121 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     9181 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     9241 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     9301 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     9361 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     9421 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     9481 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     9541 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     9601 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     9661 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
     9721 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccc cggctagccg
     9781 ccgccaccat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     9841 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     9901 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     9961 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
    10021 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
    10081 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
    10141 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
    10201 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
    10261 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
    10321 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
    10381 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
    10441 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
    10501 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
    10561 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
    10621 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
    10681 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
    10741 ccgatggctg tgtagaagtc gcgtctgcgt tcgaccaggc tgcgcgttct cgcggccata
    10801 gcaaccgacg tacggcgttg cgccctcgcc ggcagcaaga agccacggaa gtccgcccgg
    10861 agcagaaaat gcccacgcta ctgcgggttt atatagacgg tccccacggg atggggaaaa
    10921 ccaccaccac gcaactgctg gtggccctgg gttcgcgcga cgatatcgtc tacgtacccg
    10981 agccgatgac ttactggcgg gtgctggggg cttccgagac aatcgcgaac atctacacca
    11041 cacaacaccg cctcgaccag ggtgagatat cggccgggga cgcggcggtg gtaatgacaa
    11101 gcgcccagat aacaatgggc atgccttatg ccgtgaccga cgccgttctg gctcctcata
    11161 tcggggggga ggctgggagc tcacatgccc cgcccccggc cctcaccctc atcttcgacc
    11221 gccatcccat cgccgccctc ctgtgctacc cggccgcgcg gtaccttatg ggcagcatga
    11281 ccccccaggc cgtgctggcg ttcgtggccc tcatcccgcc gaccttgccc ggcaccaaca
    11341 tcgtgcttgg ggcccttccg gaggacagac acatcgaccg cctggccaaa cgccagcgcc
    11401 ccggcgagcg gctggacctg gctatgctgg ctgcgattcg ccgcgtttac gggctacttg
    11461 ccaatacggt gcggtatctg cagtgcggcg ggtcgtggcg ggaggactgg ggacagcttt
    11521 cggggacggc cgtgccgccc cagggtgccg agccccagag caacgcgggc ccacgacccc
    11581 atatcgggga cacgttattt accctgtttc gggcccccga gttgctggcc cccaacggcg
    11641 acctgtataa cgtgtttgcc tgggccttgg acgtcttggc caaacgcctc cgttccatgc
    11701 acgtctttat cctggattac gaccaatcgc ccgccggctg ccgggacgcc ctgctgcaac
    11761 ttacctccgg gatggtccag acccacgtca ccacccccgg ctccataccg acgatatgcg
    11821 acctggcgcg cacgtttgcc cgggagatgg gggaggctaa ctgaagcttc ccgggggtac
    11881 cgaattcgtc gacagatcta acttgtttat tgcagcttat aatggttaca aataaagcaa
    11941 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    12001 caaactcatc aatgtatctt atcatgtctg gatcatcaag cttaggatcc ggaaccctta
    12061 atataacttc gtataatgta tgctatacga agttattagg tccctcgaag aggttcactg
    12121 gcgcgttgga tccccgggta ccgagttggg agctcacggg gacagccccc ccccaaagcc
    12181 cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc
    12241 cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg
    12301 cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct
    12361 gcagacacct ggggggatac ggggaaaaag ctttaggctg aaagagagat ttagaatgac
    12421 agtctagtgg gagctcacgg ggacagcccc cccccaaagc ccccagggat gtaattacgt
    12481 ccctcccccg ctagggggca gcagcgagcc gcccggggct ccgctccggt ccggcgctcc
    12541 ccccgcatcc ccgagccggc agcgtgcggg gacagc
//
