LOCUS       sequence-371292-       12220 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Fluorescent reportor for lipophagy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12220)
  AUTHORS   Minami Y, Hoshino A, Higuchi Y, Hamaguchi M, Kaneko Y, Kirita Y, 
            Taminishi S, Nishiji T, Taruno A, Fukui M, Arany Z, Matoba S
  TITLE     Liver lipophagy ameliorates nonalcoholic steatohepatitis through 
            extracellular lipid secretion.
  JOURNAL   Nat Commun. 2023 Jul 13;14(1):4084. doi: 10.1038/s41467-023-39404-6.
  PUBMED    37443159
REFERENCE   2  (bases 1 to 12220)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-07-13"; volume: "14(1):4084. doi"; pages: " 
            10.1038/s41467-023-39404-6"
COMMENT     Sequence Label: pLenti EF1a GFP-mCherry-mPlin1
FEATURES             Location/Qualifiers
     source          1..12220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(131..199,200..991)
                     /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(349..368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1162..1750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1651..1670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1904..1921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2074..2079,2080..2097,2098..2104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2128
                     /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     2117..2139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2136..2152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2136..2152
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2171..2191
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2173..2191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2219..2445
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2446..2626
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2673..2798
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3291..3524
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3709..3753
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    4020..4136
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4158..5336
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4389..5327
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5284..5304
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      5352..5361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5358..5360,5361..5363,5364..6074)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5358..5360,5361..5363,5364..6074)
                     /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(5403..5424)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5664..5683)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6011..6032
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6096..6800
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(6238..6256)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6506..6525
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    8397..8985
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8450..8470)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8868..8879)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        9041..9540
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9076..9099)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     9473..9492
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             9558..10583
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             9558..10583
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             10957..11190
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    11262..11383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11299..11318)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11353..11372
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      11423..11558
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(11578..11597)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(11579..11597)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11607..11624)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11607..11623)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11616..11638)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11765..12193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11852..11871)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12062..12083
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2161 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     2221 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     2281 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     2341 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     2401 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     2461 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     2521 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2581 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     2641 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     2701 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     2761 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     2821 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     2881 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     2941 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     3001 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     3061 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     3121 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     3181 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     3241 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     3301 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     3361 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     3421 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     3481 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     3541 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     3601 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     3661 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     3721 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     3781 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     3841 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     3901 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     3961 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact
     4021 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     4081 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa aattttatcg
     4141 ataaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     4201 tccccgagaa gttgggggga ggggtcggca attgatccgg tgcctagaga aggtggcgcg
     4261 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     4321 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     4381 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
     4441 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
     4501 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
     4561 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
     4621 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
     4681 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
     4741 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     4801 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     4861 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     4921 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     4981 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     5041 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     5101 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     5161 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     5221 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     5281 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgacgta
     5341 cgtctagagt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     5401 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     5461 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     5521 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     5581 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     5641 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     5701 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     5761 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     5821 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     5881 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     5941 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     6001 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     6061 acgagctgta caagggatcc ggaggaggag gttcagtgag caagggcgag gaggataaca
     6121 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     6181 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     6241 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     6301 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     6361 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     6421 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     6481 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     6541 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     6601 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     6661 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     6721 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     6781 gcatggacga gctgtacaag tccggactca gatctcgagc tcaagctttg gttaacatgt
     6841 caatgaacaa gggcccaacc ctgctggatg gagacctccc tgagcaggag aacgtgctcc
     6901 agagagttct gcagctgcct gtggtgagcg ggacctgtga gtgcttccag aagacctaca
     6961 acagcaccaa agaagcccac cccctggtgg cctctgtgtg caatgcctat gagaagggtg
     7021 tacagggtgc cagcaacctg gctgcctgga gcatggagcc ggtggtccgt cggctgtcca
     7081 cccagttcac agctgccaat gagttggcct gcagaggcct ggaccacctg gaggaaaaga
     7141 tcccggctct tcaataccct ccagaaaaga tcgcctctga actgaagggc accatctcta
     7201 cccgccttcg aagcgccagg aacagcatca gtgtgcccat tgcaagcacc tctgacaagg
     7261 ttctgggggc cactctggcc ggctgcgagc ttgccttggg gatggccaaa gagacagcag
     7321 aatatgccgc caacacccgg gttggccgac tggcctctgg aggggctgat ctggctctgg
     7381 gaagcatcga gaaggtggta gagttcctcc tgccaccaga caaggagtca gccccttctt
     7441 ccggacggca gaggacccag aaggctccca aggccaaacc aagccttgtg aggagggtca
     7501 gcaccctggc caacactctt tctcgacaca ccatgcaaac cacagcatgg gccctgaagc
     7561 agggccactc tctggccatg tggatcccgg gtgtggcacc cctgagcagc ctggcccagt
     7621 ggggcgcatc ggcagccatg caggtggtgt cccggcggca gagtgaggtg cgggtgccct
     7681 ggctgcacaa cctggcagcc tctcaggatg agagccatga cgaccagaca gacacagagg
     7741 gagaggagac agacgacgag gaggaggaag aagagtccga ggctgaggag aacgtgctca
     7801 gagaggttac agccctgccc aacccgagag gcctcctggg tggtgtggta cacaccgtgc
     7861 agaacactct ccggaacacc atctccgcag tgacctgggc acctgcggct gtgctgggca
     7921 cggtgggaag gatcctgcac ctcacaccag cccaggctgt ctcctctacc aaagggaggg
     7981 ccatgtccct atccgatgcc ctgaagggtg ttacggataa cgtggtagac actgtggtac
     8041 actatgtgcc gcttcccagg ctgtccctga tggagcccga gagcgaattc cgagacatcg
     8101 ataacccttc agcagaggcg gagcgcaaag ggtccggggc gcggcccgcc agcccggagt
     8161 ccaccccgcg cccgggccag ccccgcggca gcttgcgcag cgtgcggggt ctcagcgcgc
     8221 cctcctgccc cggcctggac gacaaaaccg aggcgtcagc gcgtcccggc ttcctggcta
     8281 tgcccagaga gaagcctgcg cgcagagtca gcgacagctt cttccggccc agcgtcatgg
     8341 agcccatcct gggccgcgcg cagtacagcc agctgcgcaa gaagagctga gtcgacaatc
     8401 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     8461 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     8521 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     8581 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     8641 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     8701 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     8761 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     8821 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     8881 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     8941 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attctgcaga
     9001 tatccggact agtgatctct cgaggttaac gaattctacc gggtagggga ggcgcttttc
     9061 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     9121 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     9181 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     9241 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     9301 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     9361 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     9421 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     9481 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     9541 acctgcatcc cgccaccatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     9601 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     9661 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     9721 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     9781 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
     9841 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     9901 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     9961 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
    10021 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
    10081 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
    10141 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
    10201 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
    10261 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
    10321 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
    10381 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
    10441 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
    10501 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
    10561 ctcgtccgag ggcaaaggaa tagagtagat gccgaccgaa caagagctga tttcgagaac
    10621 gcctcagcca gcaactcgcg cgagcctagc aaggcaaatg cgagagaacg gccttacgct
    10681 tggtggcaca gttctcgtcc acagttcgct aagctcgctc ggctgggtcg cgggagggcc
    10741 ggtcgcagtg attcaggccc ttctggattg tgttggtccc cagggcacga ttgtcatgcc
    10801 cacgcactcg ggtgatctga ctgatcccgc agattggaga tcgccgcccg tgcctgccga
    10861 ttgggtgcag atccgtcgag ggcccggtac ctttaagacc aatgacttac aaggcagctg
    10921 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctagct cactcccaac
    10981 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
    11041 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
    11101 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
    11161 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
    11221 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
    11281 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
    11341 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
    11401 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
    11461 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
    11521 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
    11581 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
    11641 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
    11701 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
    11761 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
    11821 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
    11881 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
    11941 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
    12001 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
    12061 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
    12121 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
    12181 tttaacgcga attttaacaa aatattaacg cttacaattt
//
