LOCUS       sequence-345250-       19921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The insert is a codon optimized DNA sequence encoding VPS13D with an
            internal EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19921)
  AUTHORS   Guillen-Samander A, Leonzino M, Hanna MG, Tang N, Shen H, De Camilli
            P
  TITLE     VPS13D bridges the ER to mitochondria and peroxisomes via Miro.
  JOURNAL   J Cell Biol. 2021 May 3;220(5). pii: 212021. doi: 
            10.1083/jcb.202010004.
  PUBMED    33891013
REFERENCE   2  (bases 1 to 19921)
  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: "2021-05-3"; volume: "220(5). pii: 212021. doi"; 
            pages: " 10.1083/jcb.202010004"
COMMENT     Sequence Label: VPS13D^EGFP
FEATURES             Location/Qualifiers
     source          1..19921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      5049..5058
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5055..5057,5058..5060,5061..5771)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5055..5057,5058..5060,5061..5771)
                     /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(5100..5121)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5361..5380)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5708..5729
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    14268..14856
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(14321..14341)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14739..14750)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(14878..14897)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    14922..14970
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      15172..15600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(15259..15278)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15469..15490
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(15609..15629)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        15614..15943
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      15794..15929
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     15856..15875
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             16010..16804
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(16064..16083)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     16674..16693
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    16980..17101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(17017..17036)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     17071..17090
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(17150..17166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(17150..17166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(17163..17185)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    17174..17190
                     /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(17198..17204,17205..17222,17223..17228))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    17243..17264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(17381..17398)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(17552..18140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(17632..17651)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(18311..19102,19103..19171))
                     /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     18934..18953
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(19172..19276)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(19351..19370)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        19542..19921
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagaa
      301 tgctggaggg cctggtcgct tgggtcctga acacctacct ggggaagtat gtgaacaacc
      361 tgaacaccga tcagctgtcc gtcgctctgc tgaagggcgc cgtggagctg gagaacctgc
      421 cactgaagaa ggatgccctg aaggagctgg agctgccctt cgaggtgaag gccggcttta
      481 tcggcaaggt gaccctgcag atcccttttt acaggccaca cgtggacccc tgggtcatca
      541 gcatcagctc cctgcacctg atcggcgccc ctgagaagat ccaggacttc aatgatgaga
      601 aggagaagct gctggagaga gagaggaaga aggccctgct gcaggccctg gaggagaagt
      661 ggaagaacga ccgccagcag aagggcgagt cctactggta ttctgtgacc gccagcgtgg
      721 tgacaagaat cgtggagaat atcgagctga agatccagga cgtgcacctg aggttcgagg
      781 atggcgtgac caacccttct cacccattcg cctttggcat ctgcatcaag aatgtgagca
      841 tgcagaatgc cgtgaacgag cccgtgcaga agctgatgcg gaagaagcag ctggatgtgg
      901 ccgagttttc catctactgg gacgtggatt gtacactgct gggcgacctg cctcagatgg
      961 agctgcagga ggcaatggca cgcagcatgg agagccggtc ccaccactat gtgctggagc
     1021 cagtgttcgc cagcgccctg ctgaagcgca actgctccaa gaagcctctg cgctctcggc
     1081 acagcccacg gatcgactgt gatatccagc tggagacaat ccccctgaag ctgtcccagc
     1141 tgcagtacag acagatcatg gagtttctga aggagctgga gcgcaaggag cggcaggtga
     1201 agttccggag atggaagcct aaggtggcca tcagcaagaa ctgccgcgag tggtggtatt
     1261 tcgccctgaa cgccaatctg tatgagatcc gggagcagag aaagaggtgt acctgggact
     1321 ttatgctgca cagggcaagg gacgccgtgt cttacaccga taagtatttc aataagctga
     1381 agggcggcct gctgagcaca gacgataagg aggagatgtg ccggatcgag gaggagcagt
     1441 cctttgagga gctgaagatc ctgcgcgagc tggtgcacga tcggttccac aagcaggagg
     1501 agctggccga gtctctgaga gagccccagt ttgacagccc tggcgcctgt ccaggagcac
     1561 cagagcctgg aggaggctcc ggaatgctgc agtacctgca gtcttggttc cctggatggg
     1621 gaggatggta tggacagcag accccagagg gaaacgtggt ggagggcctg tccgccgagc
     1681 agcaggagca gtggattccc gaggagatcc tgggcaccga ggagttcttt gacccaacag
     1741 ccgatgccag ctgcatgaac acatacacca agcgcgacca cgtgttcgcc aagctgaatc
     1801 tgcagctgca gcggggcaca gtgaccctgc tgcacaagga gcagggaacc ccacagatga
     1861 acgagagcgc cttcatgcag ctggagtttt ccgatgtgaa gctgctggcc gagtctctgc
     1921 ctaggcgcaa ttctagcctg ctgagcgtgc gcctgggagg cctgtttctg cgggacctgg
     1981 caacagaggg caccatgttc cccctgctgg tgtttccaaa cccccagaag gaagtgggca
     2041 gagtgtctca gagcttcggc ctgcagacca catccgccga caggtctgat cactacccag
     2101 cagccgaccc cgatggcccc gtgttcgaga tgctgtatga gagaaatccc gcccacagcc
     2161 acttcgagcg gagactgaac gtgtccacca ggcccctgaa tatcatctac aaccctcagg
     2221 ccatcaagaa ggtggccgac ttcttttaca agggcaaggt gcacacatct ggcttcggct
     2281 atcagagcga gctggagctg agagtggcag aggcagcaag gcgccagtat aacaagctga
     2341 agatgcagac caaggccgag atcagacaga cactggacag gctgctggtg ggcgacttca
     2401 tcgaggagtc taagcggtgg accgtgcggc tggatatcag cgccccacag gtcatcttcc
     2461 ccgacgattt caagtttaag aatcctgtgc tggtggtggt ggacctgggc agaatgctgc
     2521 tgacaaatac ccaggataac agccggagaa agtccaggga cggctccgcc tctgaggaga
     2581 cacagttctc cgacgatgag tacaagacac cactggccac cccccctaat acaccacccc
     2641 ctgagtcctc tagctccaac ggcgagaaga ccccaccctt ctccggcgtg gagttttctg
     2701 aggagcagct gcaggcccac ctgatgagca caaagatgta cgagcggtac agcctgtcct
     2761 ttatggatct gcagatcatg gtgggccggg tgaaggacaa ctggaagcac gtgcaggaca
     2821 tcgatgtggg ccccacccac gtggtggaga agttcaatgt gcacctgcag ctggagaggc
     2881 gcctgatcta cacatccgac ccaaagtatc caggagccgt gctgtctgga aacctgcctg
     2941 atctgaagat ccacatcaat gaggacaaga tcagcgccct gaagaactgc tttgccctgc
     3001 tgaccacacc agagatgaag acatccgata cccagatcaa ggagaagatc ttcccacagg
     3061 aggagcagag gggctctctg caggacagcg tgatgaacct gacccagtct atcgtgctgc
     3121 tggagcagca cacaagagag gtgctggtgg agagccagct gctgctggcc gagttcaagg
     3181 tcaattgtat gcagctgggc gtggagtcca acggcagata catctctgtg ctgaaggtgt
     3241 tcggcaccaa cgcccacttt gtgaagaggc catatgatgc cgaggtgagc ctgacagtgc
     3301 acggcctgct gctggtggac acaatgcaga cctacggcgc cgactttgat ctgctgatgg
     3361 ccagccacaa gaatctgtcc ttcgatatcc ccaccggctc cctgagagac tctagggcac
     3421 agtcccccgt gtctggacct aatgtggcac acctgaccga cggagccaca ctgaacgata
     3481 ggtctgccac ctctgtgagc ctggacaaga tcctgacaaa ggagcaggag agcctgatca
     3541 agctggagta tcagttcgtg tctagcgagt gcccctctat gaacctggat agcaccctgc
     3601 aggtcatcag cctgcaggtg aacaatctgg acatcatcct gaatcctgag acaatcgtgg
     3661 agctgatcgg cttcctgcag aagtcttttc caaaggagaa ggacgatctg agccctcagc
     3721 cactgatgac cgatttcgag cggagcttcc gggagcaggg cacctaccag tctacatatg
     3781 agcagaatac cgaggtggcc gtggagatcc acagactgaa cctgctgctg ctgaggacag
     3841 tgggcatggc caatcgcgag aagtacggaa ggaagatcgc aaccgcaagc atcggcggca
     3901 caaaagtgaa tgtgagcatg ggctccacct ttgacatgaa cggctccctg ggctgcctgc
     3961 agctgatgga cctgacacag gataatgtga agaaccagta tgtggtgagc atcggcaatt
     4021 ccgtgggcta cgagaacatc atctctgata tcggctattt cgagagcgtg tttgtgcgca
     4081 tggaggacgc cgccctgacc gaggccctgt ccttcacatt tgtggagcgg tctaagcagg
     4141 agtgtttcct gaatctgaag atggccagcc tgcactacaa ccactccgcc aagtttctga
     4201 aggagctgac cctgtctatg gacgagctgg aggagaactt caggggcatg ctgaagagcg
     4261 ccgccacaaa ggtgaccaca gtgctggcca caaagaccgc cgagtactct gagatggtga
     4321 gcctgttcga gacacctaga aagacaaggg agcccttcat cctggaggag aatgagatct
     4381 atggcttcga cctggcctcc tctcacctgg ataccgtgaa gctgatcctg aacatcaata
     4441 tcgagtcccc cgtggtgtct atccctagga agccaggctc ccccgagctg ctggtgggac
     4501 acctgggaca gatcttcatc cagaactttg tggccggcga cgatgagtcc cgctctgatc
     4561 ggctgcaggt ggagatcaag gacatcaagc tgtacagcct gaattgcacc cagctggcag
     4621 gaagagaggc agtgggctct gagggcagca ggatgttttg tcctccaagc ggctccggct
     4681 ctgccaactc ccaggaggag gcacacttca cacgccacga cttctttgag tctctgcacc
     4741 ggggccaggc ctttcacatc ctgaacaata ccacaatcca gttcaagctg gagaagatcc
     4801 caatcgagcg ggagagcgag ctgaccttta gcctgtcccc cgacgatctg ggcacaagct
     4861 ccatcatgaa gatcgagggc aagttcgtga atccagtgca ggtggtgctg gccaagcacg
     4921 tgtacgagca ggtgctgcag accctggaca acctggtgta cagcgaggat ctgaataagt
     4981 atccagcatc cgccacatct agcccatgcc ctgattcccc actgccaggc tctggctcct
     5041 ctggatccgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     5101 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     5161 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     5221 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     5281 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     5341 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     5401 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     5461 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     5521 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     5581 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     5641 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     5701 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     5761 agctgtacaa gtccggactc agatctggat ccggaggacc cctgtctaca tgcggcgagt
     5821 ctagcgtgga gagaaaggag aatggcctgt tttctcactc ctctctgtcc aacacctctc
     5881 agaagtctct gagcgtgaag gaggtgaaga gcttcaccca gatccaggcc acattttgta
     5941 tcagcgagct gcaggtgcag ctgtccggcg atctgaccct gggagcacag ggcctggtgt
     6001 ctctgaagtt ccaggatttt gaggtggagt tcagcaagga ccacccccag acactgtcta
     6061 tccagatcgc cctgcacagc ctgctgatgg aggacctgct ggagaagaat cctgatagca
     6121 agtacaagaa cctgatggtg tccaggggag caccaaagcc tagctccctg gcacagaagg
     6181 agtacctgtc ccagtcttgc cctagcgtgt ccaacgtgga atatcctgac atgccacgca
     6241 gcctgccatc ccacatggag gaggccccca acgtgttcca gctgtaccag cgccctacct
     6301 ctgccagccg gaagaagcag aaggaggtgc aggacaagga ttatccactg accccacccc
     6361 ctagccctac agtggatgag ccaaagatcc tggtgggcaa gagcaagttt gacgattccc
     6421 tggtgcacat caacatcttc ctggtggaca agaagcaccc agagttttct agctcctaca
     6481 atagagtgaa caggtccatc gacgtggatt tcaattgtct ggacgtgctg atcacactgc
     6541 agacctgggt ggtcatcctg gacttcttcg gaatcggcag caccgcagac aaccacgcca
     6601 tgagactgcc acccgagggc atcctgcaca atgtgaagct ggagccccac gcctctatgg
     6661 agagcggcct gcaggaccct gtgaacacaa agctggatct gaaggtgcac tccctgtctc
     6721 tggtgctgaa taagaccaca tctgagctgg ccaaggccaa cgtgagcaag ctggtggccc
     6781 acctggagat gatcgagggc gatctggccc tgcagggctc catcggcagc ctgtccctgt
     6841 ctgacctgac ctgccacggc gagttctaca gagagaggtt taccacaagc ggcgaggagg
     6901 ccctgatctt ccagaccttc aagtatggcc ggccagatcc cctgctgagg agagagcacg
     6961 acatccgcgt gtctctgcgg atggccagcg tgcagtacgt gcacacccag agattccagg
     7021 ccgaggtggt ggccttcatc cagcacttta cacagctgca ggatgtgctg ggccgccaga
     7081 gggcagcaat cgagggacag accgtgaggg atcaggcaca gcggtgctcc cgggtgctgc
     7141 tggacatcga ggcaggagcc cccgtgctgc tgatccctga gtctagccgc agcaacaatc
     7201 tgatcgtggc caacctgggc aagctgaagg tgaagaataa gttcctgttt gccggcttcc
     7261 ccggcacctt ttctctgcag gacaaggaga gcgtgccaag cgcctcccct acaggcatcc
     7321 caaagcactc cctgagaaag accacatcta ccgaggagcc aaggggaaca cactcccagg
     7381 gccagtttac catgcctctg gccggcatgt ccctgggctc tctgaagagc gagttcgtgc
     7441 catctacaag caccaagcag cagggccctc agccaacact gagcgtggga caggagtcct
     7501 ctagcccaga ggatcacgtg tgcctgctgg actgcgtggt ggtggatctg caggacatgg
     7561 atatcttcgc cgccgagaga caccctaggg agtacagcaa ggccccagag gactcctctg
     7621 gcgatctgat cttcccctct tattttgtga ggcagaccgg cggcagcctg ctgacagagc
     7681 cttgccgcct gaagctgcag gtggagcgga acctggataa ggagatctcc cacaccgtgc
     7741 ctgacatctc tatccacggc aatctgagct ccgtgcactg ttccctggac ctgtacaagt
     7801 ataagctgat ccgcggcctg ctggagaaca atctgggcga gccaatcgag gagttcatgc
     7861 ggccctacga cctgcaggac cccagaatcc acaccgtgct gagcggcgag gtgtatacat
     7921 gcatgtgctt cctgatcgat atggtgaacg tgagcctgga gctgaaggac cccaagagga
     7981 aggagggagc aggctccctg gcacggttcg acttcaagaa gtgcaagctg ctgtacgagt
     8041 ccttttctaa tcagaccaag tccatcaacc tggtgagcca ctccatgatg gccttcgaca
     8101 cacggtatgc cggccagaag acctccccag gcatgacaaa cgtgttctct tgcatctttc
     8161 agcccgccaa gaattctagc accacacagg gctccatcca gatcgagctg cacttccgct
     8221 ctaccaagga ctcctcttgt tttacagtgg tgctgaacaa tctgcgggtg ttcctgatct
     8281 ttgactggct gctgctggtg cacgatttcc tgcacacccc aagcgacatc aagaagcaga
     8341 atcacgtgac acccagcaga cacaggaaca gctcctctga gtccgccatc gtgccaaaga
     8401 ccgtgaagtc tggcgtggtg acaaagagaa gctccctgcc cgtgtccaat gagaggcacc
     8461 tggaggtgaa ggtgaacgtg acaggcaccg agttcgtggt catcgaggac gtgagctgct
     8521 ttgataccaa cgccatcatc ctgaagggca ccacagtgct gacatacaag ccaagattcg
     8581 tggacaggcc cttttctggc agcctgttcg gcatcgaggt gtttagctgt agactgggca
     8641 acgagcacga taccgccctg tccatcgtgg accctgtgca gatccagatg gagctggtgg
     8701 gcaattctag ctaccagaac tcctctggcc tgatggacgc cttcaattcc gaggattttc
     8761 ctccagtgct ggagatccag ctgcaggccc tggatatcag gctgtcctat aacgacgtgc
     8821 agctgttcct ggccatcgcc aagtctatcc ctgagcaggc caatgcagca gtgccagata
     8881 gcgtggccct ggagtccgac tctgtgggca cctacctgcc aggagcatcc cgcgtgggag
     8941 aggagatcag ggagggaaca aggcacaccc tggaccccgt gctggagctg cagctggcaa
     9001 gactgcagga gctgggcttt tccatggacg attgcaggaa ggccctgctg gcatgtcagg
     9061 gacagctgaa gaaggccgcc agctggctgt tcaagaacgc agagcccctg aagtccctgt
     9121 ccctggcatc taccagcagg gacagccctg gagcagtggc agcaccactg atcagcggcg
     9181 tggagatcaa ggccgagtcc gtgtgcatct gtttcatcga cgattgcatg gactgtgatg
     9241 tgcctctggc cgagctgaca ttctccaggc tgaactttct gcagcgcgtg cggacctctc
     9301 cagagggata cgcccacttt acactgagcg gcgactacta taatcgcgcc ctgtccggct
     9361 gggagccatt catcgagccc tggccttgct ccgtgtcttg gcagcagcag gcagcatcca
     9421 gactgcaccc acctaggctg aagctggagg caaaggcaaa gccacgcctg gatatcaaca
     9481 tcaccagcgt gctgatcgac cagtacgtga gcacaaagga gtcctggatg gccgattatt
     9541 gcaaggacga taaggacatc gagtccgcca agtctgagga ttggatgggc agctccgtgg
     9601 acccaccctg tttcggccag agcctgcccc tggtgtacct gagaacaagg agcaccgcct
     9661 ccctgaccaa cctggagcac cagatctatg cccgcgccga ggtgaagaca cctaagaggc
     9721 gccagccctt cgtgcctttt gccctgcgga atcacacagg ctgtaccctg tggtttgcca
     9781 ccctgaccac aaccccaaca agagccgccc tgagccactc cggctctccc ggagtggtgc
     9841 ctgagggaaa tggcaccttc ctggacgata cacacaacgt gtccgagtgg agggaggtgc
     9901 tgaccggcga ggagatcccc ttcgagtttg aggcaagggg caagctgagg cacaggcaca
     9961 cacacgacct gagaatccac cagctgcagg tgagggtgaa cggctgggag caggtgtctc
    10021 ctgtgagcgt ggataaagtg ggcaccttct ttcggtacgc cgccccagac aagaactcta
    10081 gctcctctac catcggcagc cccagctcca gaacaaatat catccaccct caggtgtatt
    10141 tttctagcct gcctccagtg agggtggtgt tcgccgtgac catggagggc tctgccagaa
    10201 aagtgatcac agtgaggagc gccctgatcg tgagaaaccg gctggagaca cccatggagc
    10261 tgagactgga cagcccatcc gcccccgata agcctgtggt gctgccagcc atcatgcccg
    10321 gcgattcctt tgccgtgcct ctgcacctga catcttggag actgcaggcc aggcctaagg
    10381 gcctgggcgt gttcttttgc aaggccccaa tccactggac caatgtggtg aagacagccg
    10441 agatctcctc tagcaagagg gagtgccaca gcatggacac tgagaagtcc cgcttctttc
    10501 ggttctgcgt ggccatcaag aaggagaact accctgatta tatgccaagc aatatcttct
    10561 ctgacagcgc caagcagatc tttcgccagc ccggccacac catctacctg ctgcctacag
    10621 tggtcatctg caacctgctg ccctgtgagc tggacttcta cgtgaagggc atgcccatca
    10681 atggcaccct gaagcctggc aaggaggccg ccctgcacac agcagacacc tctcagaata
    10741 tcgagctggg cgtgagcctg gagaacttcc ctctgtgcaa ggagctgctg atcccccctg
    10801 gcacccagaa ctacatggtg cgcatgcggc tgtatgacgt gaatcggaga cagctgaacc
    10861 tgacaatcag gatcgtgtgc agggcagagg gctctctgaa gatcttcatc agcgcccctt
    10921 actggctgat caacaagacc ggcctgccac tgatctttag gcaggacaat gccaagacag
    10981 atgcagcagg acagttcgag gagcacgagc tggcacgctc cctgtctccc ctgctgtttt
    11041 gctatgccga taaggagcag cctaacctgt gcaccatgag aatcggaagg ggcatccacc
    11101 cagagggaat gcctggatgg tgccagggct tctccctgga cggaggctct ggcgtgcggg
    11161 ccctgaaagt gatccagcag ggcaataggc ctggcctgat ctacaacatc ggcatcgatg
    11221 tgaagaaggg cagaggcagg tacatcgaca cctgtatggt catctttgcc ccacggtatc
    11281 tgctggacaa caagtcctct cacaagctgg ccttcgccca gagagagttt gcaaggggac
    11341 agggaaccgc aaatccagag ggctacatca gcacactgcc aggcagctcc gtggtgttcc
    11401 actggccccg caacgactat gatcagctgc tgtgcgtgcg gctgatggat gtgcctaatt
    11461 gtatctggag cggcggcttc gaggtgaaca agaacaattc ctttcacatc aatatgagag
    11521 acaccctggg caagtgcttc tttctgaggg tggagatcac cctgagggga gcaacatacc
    11581 ggatcagctt ttccgacacc gatcagctgc cacccccttt caggatcgac aacttttcca
    11641 aggtgcctgt ggtgttcaca cagcacggag tggcagagcc taggctgagg accgaggtga
    11701 agccaatgac aagcctggac tatgcctggg atgagccaac cctgccaccc tttatcacac
    11761 tgaccgtgaa gggcgccggc tctagcgaga tcaactgcaa tatgaacgac ttccaggata
    11821 atcgccagct gtactatgag aacttcatct acatcgccgc cacatatacc ttttccggcc
    11881 tgcaggaggg aaccggccgg cccgtggcct ctaacaaggc catcacatgt gccgagctgg
    11941 tgctggacgt gagccctaag acccagagag tgatcctgaa gaagaaggag ccaggcaaga
    12001 gatctcagct gtggaggatg acaggcaccg gcatgctggc acacgagggc tcctctgtgc
    12061 cacacaatcc caacaagcct tctgccgcac ggagcaccga gggctccgcc atcctggata
    12121 tcgcaggcct ggcagcagtg acagacaatc gctacgagcc cctgatgctg aggaagcctg
    12181 acaggcgccg gtccacaacc cagacctggt cttttagaga gggcaagctg acatgcggcc
    12241 tgcacggcct ggtggtgcag gcaaagggag gcctgtccgg cctgttcgat ggagcagagg
    12301 tggtgctggg accagacacc tctatggagc tgctgggccc cgtgcctcca gagcagcagt
    12361 ttatcaacca gaagatgcgg ccaggctctg gcatgctgag catccgcgtg atcccagatg
    12421 gaccaacccg ggccctgcag atcacagact tctgtcaccg caagagctcc cggtcctatg
    12481 aggtggatga gctgcccgtg accgagcagg agctgcagaa gctgaagaat cctgacacag
    12541 agcaggagct ggaggtgctg gtgagactgg agggcggcat tggcctgtcc ctgatcaaca
    12601 aggtgcccga ggagctggtg tttgcctctc tgaccggcat caatgtgcac tacacccagc
    12661 tggccacaag ccacatgctg gagctgtcca tccaggacgt gcaggtggat aaccagctga
    12721 tcggcacaac ccagccattc atgctgtatg tgacccccct gagcaatgag aacgaagtga
    12781 tcgagacagg cccagcagtg caggtgaacg cagtgaagtt tccttctaag agcgccctga
    12841 ccaatatcta caagcacctg atgatcaccg cccagcggtt cacagtgcag atcgaggaga
    12901 agctgctgct gaagctgctg agcttctttg gctacgatca ggccgagtcc gaggtggaga
    12961 agtatgacga gaatctgcac gagaagaccg cagagcaggg aggaacacct atcagatact
    13021 attttgagaa cctgaagatc tccatcccac agatcaagct gtccgtgttc acctctaata
    13081 agctgcccct ggatctgaag gccctgaaga gcacactggg cttccctctg atcagatttg
    13141 aggacgccgt gatcaacctg gaccccttca ccagggtgca cccctacgag acaaaggagt
    13201 tcatcatcaa tgacatcctg aagcactttc aggaggagct gctgtcccag gccgccagaa
    13261 tcctgggctc tgtggatttc ctgggcaatc ctatgggcct gctgaacgac gtgtccgagg
    13321 gcgtgaccgg cctgatcaag tatggcaatg tgggcggcct gatccggaac gtgacacacg
    13381 gcgtgtccaa ttctgccgca aagttcgcag gcaccctgag cgatggcctg ggcaagacaa
    13441 tggacaacag acaccagagc gagagagagt acatcaggta tcacgcagcc acctccggag
    13501 agcacctggt ggccggcatc cacggcctgg cacacggaat catcggaggc ctgaccagcg
    13561 tgatcacatc caccgtggag ggagtgaaga cagagggagg cgtgtctggc tttatcagcg
    13621 gcctgggcaa gggcctggtg ggaacagtga ccaagcccgt ggcaggcgcc ctggatttcg
    13681 caagcgagac agcacaggcc gtgcgcgaca cagccaccct gtccggacca aggacccagg
    13741 cacagagagt gaggaagcca aggtgctgta caggcccaca gggcctgctg ccaaggtaca
    13801 gcgagtccca ggcagaggga caggagcagc tgttcaagct gaccgacaac atccaggatg
    13861 agttctttat cgccgtggag aatatcgata gctattgcgt gctgatctct agcaaggccg
    13921 tgtactttct gaaatccggc gactatgtgg atcgggaggc catcttcctg gaggtgaagt
    13981 acgacgatct gtatcactgt ctggtgtcca aggaccacgg caaggtgtac gtgcaggtga
    14041 ccaagaaggc cgtgtctaca tcctctggcg tgagcatccc aggaccatcc caccagaagc
    14101 ccatggtgca cgtgaagagc gaggtgctgg ccgtgaagct gtcccaggag atcaactatg
    14161 ccaagtctct gtactatgaa cagcagctga tgctgagact gagcgaaaac agggaacagc
    14221 tggaactgga ttcataaacc ggttagtaat gagtttgata tctcgacaat caacctctgg
    14281 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
    14341 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
    14401 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
    14461 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
    14521 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
    14581 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
    14641 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
    14701 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
    14761 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
    14821 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga
    14881 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
    14941 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
    15001 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
    15061 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
    15121 caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct
    15181 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
    15241 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
    15301 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac
    15361 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
    15421 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
    15481 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
    15541 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
    15601 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
    15661 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
    15721 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
    15781 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
    15841 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
    15901 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
    15961 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
    16021 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
    16081 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
    16141 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
    16201 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
    16261 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
    16321 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
    16381 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
    16441 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
    16501 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
    16561 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
    16621 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
    16681 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
    16741 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
    16801 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac
    16861 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
    16921 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
    16981 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
    17041 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
    17101 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
    17161 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    17221 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    17281 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    17341 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
    17401 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
    17461 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
    17521 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
    17581 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
    17641 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
    17701 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
    17761 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
    17821 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
    17881 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    17941 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
    18001 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    18061 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    18121 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    18181 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    18241 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    18301 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    18361 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    18421 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    18481 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    18541 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    18601 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    18661 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    18721 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    18781 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    18841 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    18901 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    18961 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    19021 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    19081 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    19141 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    19201 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    19261 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
    19321 atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag
    19381 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
    19441 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
    19501 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
    19561 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
    19621 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
    19681 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
    19741 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
    19801 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
    19861 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
    19921 g
//
