LOCUS       sequence-346521-       12576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac Vector expressing GFP-RFP-LGALS3 .
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
FEATURES             Location/Qualifiers
     source          1..12576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    325..549
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     538..555
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(593..634)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    687..711
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1475..2185,2186..2188,2189..2191))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1475..2185,2186..2188,2189..2191))
                     /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     1866..1885
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2125..2146
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2188..2197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2210..2884)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             complement(2210..2884)
                     /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(2269..2292)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2467..2486)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2736..2754
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(2884..2908)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2969..4147)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2978..3916
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(3001..3021)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(4789..4822)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4851..4985
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4875..4894)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4929..4948
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5021..6016)
                     /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"
     primer_bind     complement(7180..7204)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(7201..7404)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7230..7250)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7405..7784
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    complement(7934..7967)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   8301..8335
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(8374..8392)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8413..8429)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8413..8429)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8426..8448)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8437..8453
                     /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(8461..8467,8468..8485,8486..8491))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8506..8527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8644..8661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8815..9403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8895..8914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(9527..9595,9596..10387)
                     /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(9745..9764)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(10637..11431)
                     /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             complement(10637..11431)
                     /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(10748..10767)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     11358..11377
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(11780..12082)
                     /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     complement(11882..11901)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     12205..12227
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12219..12236
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12220..12236
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12243..12262
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        12243..12261
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   12300..12362
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 tctgcaatga aatcgagttc tgtcattcta aatctctctt tcagcctaaa gctttttccc
       61 cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga
      121 tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg
      181 ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag
      241 ggacgtaatt acatccctgg gggctttggg ggggggctgt ccccgtgagc tcccaactcg
      301 gtacccgggg atccaggcgc gccgccatag agcccaccgc atccccagca tgcctgctat
      361 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
      421 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
      481 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
      541 aaggcacagt cgaggctgat cagcgggttt aaacgggccc ctaaccggta cgcgtagaat
      601 cgagaccgag gagagggtta gggataggct taccttcgaa ccgcgggccc tctagactcg
      661 agcggccgcc actgtgctgg atatcaacca ctttgtacaa gaaagttggt atcatggtat
      721 atgaagcact ggtgaggtct atgtcaccag aaattcccag tttgctgatt tcattgagtt
      781 ttttaacccg atgattgtac tgcaacaagt gagcatcatt cactgcaacc ttgaagtggt
      841 caggttcaac cagtacttgt attttgaatg gtttcccact ttcaaatggg aaaaccgact
      901 gtctttcttc ccttccccag ttattatcca gctttgtatt gcaaacaatg actctcctgt
      961 tgttctcatt gaagcgtggg ttaaagtgga aggcaacatc attccctctt tggaaatcta
     1021 aagcaattct gtttgcattg ggcttcaccg tgcccagaat tgttatcagc atgcgaggca
     1081 ccactccccc aggcaaaggc aggttataag gcacaatcag tggcccagca ggggcgccat
     1141 aggggccagt ggcagggtag gctccggggg cacttggctg tccagaagat gggtaggccc
     1201 cagggccgct gggtggccct gggtagactc caggtgcagg tgctccggga taagctccag
     1261 gtgctccatg gtaggcgcct ggaggtgcct gtccaggata agcccctggg ggtgcctgcc
     1321 cggggtaggc cccaggatag gaagcccctg ggtagccccc tgccccagca ggctggttcc
     1381 cccatgcgcc aggccatcct tgagggtttg ggtttccaga cccagataac gcatcatgga
     1441 gcgaaaaatt gtctgccata gatctgagtc cggacttgta cagctcgtcc atgccgagag
     1501 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     1561 ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     1621 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     1681 tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga
     1741 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     1801 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     1861 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     1921 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     1981 actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     2041 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     2101 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     2161 cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtccaccg gcgccggtgg
     2221 agtggcggcc ctcggcgcgc tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg
     2281 tgatgtccag cttgatgtcg gtcttgtagg cgccgggcag ctgcacgggc ttcttggcca
     2341 tgtaggtggt cttgacctcg gcgtcgtagt ggccgccgtc cttcagcttc agcctcatct
     2401 tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggtggag gcctcccagc
     2461 ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca
     2521 ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc
     2581 cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt
     2641 agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtactgg aactgagggg
     2701 acaggatgtc ccaggcgaag ggcagggggc cgcccttggt caccttcagc ttggcggtct
     2761 gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca
     2821 cggagccctc catgcgcacc ttgaagcgca tgaactcctt gatgacgtcc tcggaggagg
     2881 ccatgccaac ttttttgtac aaacttgttg atatctgcag aattccacca cactggacta
     2941 gtggatccga gctcggtacc aagcttcttc acgacacctg aaatggaaga aaaaaacttt
     3001 gaaccactgt ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca
     3061 aggagaatta catcaagtgc caagctggcc taacttcagt ctccacccac tcagtgtggg
     3121 gaaactccat cgcataaaac ccctcccccc aacctaaaga cgacgtactc caaaagctcg
     3181 agaactaatc gaggtgcctg gacggcgccc ggtactccgt ggagtcacat gaagcgacgg
     3241 ctgaggacgg aaaggccctt ttcctttgtg tgggtgactc acccgcccgc tctcccgagc
     3301 gccgcgtcct ccattttgag ctccctgcag cagggccggg aagcggccat ctttccgctc
     3361 acgcaactgg tgccgaccgg gccagccttg ccgcccaggg cggggcgata cacggcggcg
     3421 cgaggccagg caccagagca ggccggccag cttgagacta cccccgtccg attctcggtg
     3481 gccgcgctcg caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg gccccgtcgc
     3541 cgcccgcggc cccaaaaacc gaaataccag tgtgcagatc ttggcccgca tttacaagac
     3601 tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg gctagagact
     3661 tatcgaaagc agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc
     3721 gcccaggcca ggcctcaact caagcacgag gcgaaggggc tccttaagcg caaggcctcg
     3781 aactctccca cccacttcca acccgaagct cgggatcaag aatcacgtac tgcagccagg
     3841 tggaagtaat tcaaggcacg caagggccat aacccgtaaa gaggccaggc ccgcgggaac
     3901 cacacacggc acttacctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg
     3961 cgactactgc acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca
     4021 cgacatcact ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga
     4081 cccctccccc caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac
     4141 cggagcctca cgggcgcgcc agtctgcaat gaaatcgagt tctgtcattc taaatctctc
     4201 tttcagccta aagctttttc cccgtatccc cccaggtgtc tgcaggctca aagagcagcg
     4261 agaagcgttc agaggaaagc gatcccgtgc caccttcccc gtgcccgggc tgtccccgca
     4321 cgctgccggc tcggggatgc ggggggagcg ccggaccgga gcggagcccc gggcggctcg
     4381 ctgctgcccc ctagcggggg agggacgtaa ttacatccct gggggctttg ggggggggct
     4441 gtccccgtga gctcccacta gactgtcatt ctaaatctct ctttcagcct aaagcttttt
     4501 ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc gagaagcgtt cagaggaaag
     4561 cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc acgctgccgg ctcggggatg
     4621 cggggggagc gccggaccgg agcggagccc cgggcggctc gctgctgccc cctagcgggg
     4681 gagggacgta attacatccc tgggggcttt gggggggggc tgtccccgtg agctcccaac
     4741 tcggtacccg gggatccaac gcgccagtga acctcttcga gggacctaat aacttcgtat
     4801 agcatacatt atacgaagtt atattaaggg ttccggatcc taagcttgat gatccagaca
     4861 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     4921 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     4981 aagttagatc tgtcgacgaa ttcggtaccc ccgggaagct tcagttagcc tcccccatct
     5041 cccgggcaaa cgtgcgcgcc aggtcgcata tcgtcggtat ggagccgggg gtggtgacgt
     5101 gggtctggac catcccggag gtaagttgca gcagggcgtc ccggcagccg gcgggcgatt
     5161 ggtcgtaatc caggataaag acgtgcatgg aacggaggcg tttggccaag acgtccaagg
     5221 cccaggcaaa cacgttatac aggtcgccgt tgggggccag caactcgggg gcccgaaaca
     5281 gggtaaataa cgtgtccccg atatggggtc gtgggcccgc gttgctctgg ggctcggcac
     5341 cctggggcgg cacggccgtc cccgaaagct gtccccagtc ctcccgccac gacccgccgc
     5401 actgcagata ccgcaccgta ttggcaagta gcccgtaaac gcggcgaatc gcagccagca
     5461 tagccaggtc cagccgctcg ccggggcgct ggcgtttggc caggcggtcg atgtgtctgt
     5521 cctccggaag ggccccaagc acgatgttgg tgccgggcaa ggtcggcggg atgagggcca
     5581 cgaacgccag cacggcctgg ggggtcatgc tgcccataag gtaccgcgcg gccgggtagc
     5641 acaggagggc ggcgatggga tggcggtcga agatgagggt gagggccggg ggcggggcat
     5701 gtgagctccc agcctccccc ccgatatgag gagccagaac ggcgtcggtc acggcataag
     5761 gcatgcccat tgttatctgg gcgcttgtca ttaccaccgc cgcgtccccg gccgatatct
     5821 caccctggtc gaggcggtgt tgtgtggtgt agatgttcgc gattgtctcg gaagccccca
     5881 gcacccgcca gtaagtcatc ggctcgggta cgtagacgat atcgtcgcgc gaacccaggg
     5941 ccaccagcag ttgcgtggtg gtggttttcc ccatcccgtg gggaccgtct atataaaccc
     6001 gcagtagcgt gggcattttc tgctccgggc ggacttccgt ggcttcttgc tgccggcgag
     6061 ggcgcaacgc cgtacgtcgg ttgctatggc cgcgagaacg cgcagcctgg tcgaacgcag
     6121 acgcgacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac
     6181 gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg
     6241 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca
     6301 tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg
     6361 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt
     6421 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg
     6481 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct
     6541 cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca
     6601 tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg
     6661 tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat
     6721 cggccgcagc gatcgcatcc atggcctccg cgaccggctg cagaacagcg ggcagttcgg
     6781 tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc
     6841 tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt
     6901 gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat
     6961 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca
     7021 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga
     7081 gttcaggctt tttcatggtg gcggcggcta gccggggagg ctggatcggt cccggtgtct
     7141 tctatggagg tcaaaacagc gtggatggcg tctccaggcg atctgacggt tcactaaacg
     7201 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
     7261 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     7321 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     7381 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     7441 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     7501 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     7561 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     7621 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     7681 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     7741 ccccgtaatt gattactatt aataactagt caataatcaa tgtcaacatg gcggtaatgt
     7801 tggacatgag ccaatataaa tgtacatatt atgatatgga tacaacgtat gcaatgggcc
     7861 aagctcctcg aggcccggcc atcggtaccc gggatcctaa gcttggctgg acgtaaactc
     7921 ctcttcagac ctaataactt cgtatagcat acattatacg aagttatatt aagggttatt
     7981 gaatatgatc ggaattcttg aagacgaaag gtctagcatt ctagttgata tctataacaa
     8041 gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa tataattatc
     8101 gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg
     8161 gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct
     8221 ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag
     8281 agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaaa
     8341 agggcgaatt cgtttaaacc tgcaggacta gtccctttag tgagggttaa ttctgagctt
     8401 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     8461 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     8521 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     8581 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     8641 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     8701 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     8761 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     8821 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8881 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8941 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     9001 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     9061 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     9121 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     9181 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     9241 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     9301 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     9361 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     9421 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     9481 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     9541 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     9601 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     9661 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     9721 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     9781 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     9841 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     9901 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     9961 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
    10021 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
    10081 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
    10141 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
    10201 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
    10261 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
    10321 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
    10381 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
    10441 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgagcg gatacatatt
    10501 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
    10561 acctgatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
    10621 tgtaagcgtt aataattcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
    10681 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
    10741 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
    10801 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
    10861 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
    10921 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
    10981 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
    11041 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
    11101 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
    11161 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
    11221 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
    11281 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
    11341 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
    11401 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
    11461 cagagcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
    11521 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg
    11581 tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc
    11641 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
    11701 agcaccgttt ctgcggactg gctttctacg tgaaaaggat ctaggtgaag atcctttttg
    11761 ataatctcat gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
    11821 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg
    11881 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
    11941 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
    12001 ccggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
    12061 tataggtgta aaccttaaac tgccgtacgt ataggctgcg caactgttgg gaagggcgat
    12121 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
    12181 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
    12241 tgtaatacga ctcactatag ggcgaattga atttagcggc cgcgaattcg cccttttaac
    12301 cctagaaaga taatcatatt gtgacgtacg ttaaagataa tcatgcgtaa aattgacgca
    12361 tgtgttttat cggtctgtat atcgaggttt atttattaat ttgaatagat attaagtttt
    12421 attatattta cacttacata ctaataataa attcaacaaa caatttattt atgtttattt
    12481 atttattaaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa acttttaact
    12541 agtatgtcga gccggcgcgt actagtggcc ggccag
//
