LOCUS       sequence-466621-       17197 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of mouse mutant Etfdh (AAA6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17197)
  AUTHORS   Papadopoli D, Palia R, Jovanovic P, Tabaries S, Ciccolini E, 
            Sabourin V, Igelmann S, McLaughlan S, Zhan L, Kim H, Chekkal N, 
            Szkop KJ, Bertomeu T, Zeng J, Vassalakis J, Afzali F, Mzoughi S, 
            Guccione E, Tyers M, Avizonis D, Larsson O, Postovit LM, Djuranovic 
            S, Ursini-Siegel J, Siegel PM, Pollak M, Topisirovic I
  TITLE     Mitochondrial ETF insufficiency drives neoplastic growth by 
            selectively optimizing cancer bioenergetics
  JOURNAL   bioRxiv 2024.10.25.620155
REFERENCE   2  (bases 1 to 17197)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.10.25.620155"
COMMENT     Sequence Label: pLenti-mEtfdh-AAA6
FEATURES             Location/Qualifiers
     source          1..17197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             871..1134
                     /codon_start=1
                     /gene="V"
                     /product="pV"
                     /label=M13 gene V
                     /note="replication"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             1136..1237
                     /codon_start=1
                     /gene="VII"
                     /product="pVII"
                     /label=M13 gene VII
                     /note="minor coat protein"
                     /translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
     CDS             1234..1332
                     /codon_start=1
                     /gene="IX"
                     /product="pIX"
                     /label=M13 gene IX
                     /note="minor coat protein"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
     CDS             1329..1550
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFDSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     terminator      1554..1602
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             1607..2881
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TNPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQPSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     CDS             2884..3222
                     /codon_start=1
                     /gene="VI"
                     /product="pVI"
                     /label=M13 gene VI
                     /note="minor coat protein"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
     CDS             3224..4270
                     /codon_start=1
                     /gene="I"
                     /product="pI"
                     /label=M13 gene I
                     /note="phage assembly"
                     /translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
                     VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
                     QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
                     VTGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
                     TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
                     SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSLTYIDLCTVSIKKGN
                     SNEIVKCN"
     CDS             4248..5528
                     /codon_start=1
                     /gene="IV"
                     /product="pIV"
                     /label=M13 gene IV
                     /note="phage assembly"
                     /translation="MKLLNVINFVFLMFVSSSSFAQVIEMNNSPLRDFVTWYSKQTGES
                     VIVSPDVKGTVTVYSSDVKPENLRNFFISVLRANNFDMVGSNPSIIQKYNPNNQDYIDE
                     LPSSDNQEYDDNSAPSGGFFVPQNDNVTQTFKINNVRAKDLIRVVELFVKSNTSKSSNV
                     LSIDGSNLLVVSAPKDILDNLPQFLSTVDLPTDQILIEGLIFEVQQGDALDFSFAAGSQ
                     RGTVAGGVNTDRLTSVLSSAGGSFGIFNGDVLGLSVRALKTNSHSKILSVPRILTLSGQ
                     KGSISVGQNVPFITGRVTGESANVNNPFQTIERQNVGISMSVFPVAMAGGNIVLDITSK
                     ADSLSSSTQASDVITNQRSIATTVNLRDGQTLLLGGLTDYKNTSQDSGVPFLSKIPLIG
                     LLFSSRSDSNEESTLYVLVKATIVRAL"
     rep_origin      5516..5971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5997..6101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6102..6170,6171..6962)
                     /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(6320..6339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7133..7721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7622..7641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7875..7892
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8009..8030
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8045..8050,8051..8068,8069..8075)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8083..8099
                     /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     8088..8110
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8107..8123
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8107..8123
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8142..8162
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8144..8162
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8190..8416
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8417..8597
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8644..8769
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9262..9495
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9680..9724
                     /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    9990..10107
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    10125..10145
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        10146..11324
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          10377..11315
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     11272..11292
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    11324..11348
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(13224..13248)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    13266..13854
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(13319..13339)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13737..13748)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        13885..14384
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(13920..13943)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     14317..14336
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             14401..15426
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             14401..15426
                     /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             15498..15731
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    15803..15924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(15840..15859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     15894..15913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      15964..16099
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(16119..16138)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(16120..16138)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(16148..16165)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(16148..16164)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16157..16179)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      16306..16819
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             join(16824..17197,1..859)
                     /codon_start=1
                     /gene="II"
                     /product="pII"
                     /label=M13 gene II
                     /note="replication"
                     /translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSAR
                     TVEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
                     FGTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHL
                     RNVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQ
                     KEQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIF
                     DLWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRL
                     VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
                     DWYNEPVLKIA"
ORIGIN
        1 gccttgcctg tatgatttat tggatgttaa cgctactact attagtagaa ttgatgccac
       61 cttttcagct cgcgccccaa atgaaaatat agctaaacag gttattgacc atttgcgaaa
      121 tgtatctaat ggtcaaacta aatctactcg ttcgcagaat tgggaatcaa ctgttacatg
      181 gaatgaaact tccagacacc gtactttagt tgcatattta aaacatgttg agctacagca
      241 ccagattcag caattaagct ctaagccatc cgcaaaaatg acctcttatc aaaaggagca
      301 attaaaggta ctctctaatc ctgacctgtt ggagtttgct tccggtctgg ttcgctttga
      361 agctcgaatt aaaacgcgat atttgaagtc tttcgggctt cctcttaatc tttttgatgc
      421 aatccgcttt gcttctgact ataatagtca gggtaaagac ctgatttttg atttatggtc
      481 attctcgttt tctgaactgt ttaaagcatt tgagggggat tcaatgaata tttatgacga
      541 ttccgcagta ttggacgcta tccagtctaa acattttact attaccccct ctggcaaaac
      601 ttcttttgca aaagcctctc gctattttgg tttttatcgt cgtctggtaa acgagggtta
      661 tgatagtgtt gctcttacta tgcctcgtaa ttccttttgg cgttatgtat ctgcattagt
      721 tgaatgtggt attcctaaat ctcaactgat gaatctttct acctgtaata atgttgttcc
      781 gttagttcgt tttattaacg tagatttttc ttcccaacgt cctgactggt ataatgagcc
      841 agttcttaaa atcgcataag gtaattcaca atgattaaag ttgaaattaa accatctcaa
      901 gcgcaattca ctacccgttc tggtgtttct cgtcagggca agccttattc actgaatgag
      961 cagctttgtt acgttgattt gggtaatgaa tatccggtgc ttgtcaagat tactcttgat
     1021 gaaggtcagc cagcctatgc gcctggtctg tacaccgttc atctgtcctc gttcaaagtt
     1081 ggtcagttcg gttcccttat gattgaccgt ctgcgcctcg ttccggctaa gtaatatgga
     1141 gcaggtcgcg gatttcgaca caatttatca ggcgatgata caaatctccg ttgtactttg
     1201 tttcgcgctt ggtataatcg ctgggggtca aagatgagtg ttttagtgta ttctttcgcc
     1261 tctttcgttt taggttggtg ccttcgtagt ggcattacgt attttacccg tttaatggag
     1321 acttcctcat gaaaaagtct ttagtcctca aagcctctgt agccgttgct accctcgttc
     1381 cgatgctgtc tttcgctgct gagggtgacg atcccgcaaa agcggccttt gactccctgc
     1441 aagcctcagc gaccgaatat atcggttatg cgtgggcgat ggttgttgtc attgtcggcg
     1501 caactatcgg tatcaagctg tttaagaaat tcacctcgaa agcaagctga taaaccgata
     1561 caattaaagg ctccttttgg agcctttttt tttggagatt ttcaacgtga aaaaattatt
     1621 attcgcaatt cctttagttg ttcctttcta ttctcactcc gctgaaactg ttgaaagttg
     1681 tttagcaaaa cctcatacag aaaattcatt tactaacgtc tggaaagacg acaaaacatt
     1741 agatcgctac gctaactatg agggctgtct gtggaatgct acaggcgttg tagtttgtac
     1801 tggcgacgaa actcagtgtt atggtacatg ggttcctatt gggcttgcta tccctgaaaa
     1861 tgagggtggc ggctctgagg gtggcggttc tgagggtggc ggctctgagg gtggcggcac
     1921 taatcctcct gagtacggtg atacccctat tccgggctat acctatatca atcctctcga
     1981 cggcacttat ccgcctggta ctgagcaaaa ccccgctaat cctaatcctt ctcttgagga
     2041 gtctcagcct cttaatactt tcatgtttca gaataatagg ttccgaaata ggcagggtgc
     2101 attaaccgta tatacgggaa ccgttactca aggcactgac cccgtcaaaa cttattacca
     2161 gtacactcct gtttcatcaa aagccatgta cgacgcttac tggaacggta aattcagaga
     2221 ctgtgctttc cattccggct ttaatgagga tccattcgtt tgtgaatatc aaggccaacc
     2281 gtctgacctg cctcaacctc ccgttaatgc tggcggcggc tctggtggtg gttctggtgg
     2341 cggctctgag ggtggcggct ctgagggtgg cggttctgag ggtggcggct ctgagggtgg
     2401 cggctccggt ggcggctccg gttccggtga ttttgattat gaaaaaatgg caaacgctaa
     2461 taagggggct atgaccgaaa atgccgatga aaatgcgcta cagtctgacg ctaaaggcaa
     2521 acttgattct gtcgctactg attacggtgc tgctatcgac ggtttcattg gtgacgtttc
     2581 cggccttgct aatggtaatg gtgctactgg tgattttgct ggctctaatt cccaaatggc
     2641 tcaagtcggt gacggtgata attcaccttt aatgaataat ttccgtcaat atttaccttc
     2701 cctccctcaa tcggttgaat gtcgcccttt tgtctttggc gctggtaaac catatgaatt
     2761 ttctattgat tgtgacaaaa taaacttatt ccgtggtgtc tttgcgtttc ttttatatgt
     2821 tgccaccttt atgtatgtat tttcgacgtt tgctaacata ctgcgtaata aggagtctta
     2881 atcatgccag ttcttttggg tattccgtta ttattgcgtt tcctcggttt ccttctggta
     2941 actttgttcg gctatctgct tacttttctt aaaaagggct tcggtaagat agctattgct
     3001 atttcattgt ttcttgctct tattattggg cttaactcaa ttcttgtggg ttatctctct
     3061 gatattagtg ctcaattacc ctctgacttt gttcagggcg ttcagttaat tctcccgtct
     3121 aatgcgcttc cctgttttta tgttattctc tctgtaaagg ctgctatttt catttttgac
     3181 gttaaacaaa aaatcgtttc ttatttggat tgggataaat aatatggctg tttattttgt
     3241 aactggcaaa ttaggctctg gaaagacgct cgttagcgtt ggtaagattc aggataaaat
     3301 tgtagctggg tgcaaaatag caactaatct tgatttaagg cttcaaaacc tcccgcaagt
     3361 cgggaggttc gctaaaacgc ctcgcgttct tagaataccg gataagcctt ctatatctga
     3421 tttgcttgct attgggcgcg gtaatgattc ctacgatgaa aataaaaacg gcttgcttgt
     3481 tctcgatgag tgcggtactt ggtttaatac ccgttcttgg aatgataagg aaagacagcc
     3541 gattattgat tggtttctac atgctcgtaa attaggatgg gatattattt ttcttgttca
     3601 ggacttatct attgttgata aacaggcgcg ttctgcatta gctgaacatg ttgtttattg
     3661 tcgtcgtctg gacagaatta ctttgccttt tgtcggtact ttatattctc ttgttactgg
     3721 ctcgaaaatg cctctgccta aattacatgt tggcgttgtt aaatatggcg attctcaatt
     3781 aagccctact gttgagcgtt ggctttatac tggtaagaat ttgtataacg catatgatac
     3841 taaacaggct ttttccagta attatgattc cggtgtttat tcttatttaa cgccttattt
     3901 atcacacggt cggtatttca aaccattaaa tttaggtcag aaaatgaaat taactaaaat
     3961 atatttgaaa aagttttctc gcgttctttg tcttgcgatt ggatttgcat cagcatttac
     4021 atatagttat ataacccaac ctaagccgga ggttaaaaag gtagtctctc agacctatga
     4081 ttttgataaa ttcactattg actcttctca gcgtcttaat ctaagctatc gctatgtttt
     4141 caaggattct aagggaaaat taattaatag cgacgattta cagaagcaag gttattcact
     4201 cacatatatt gatttatgta ctgtttccat taaaaaaggt aattcaaatg aaattgttaa
     4261 atgtaattaa ttttgttttc ttgatgtttg tttcatcatc ttcttttgct caagtaattg
     4321 aaatgaataa ttcgcctctg cgcgatttcg ttacttggta ttcaaagcaa acaggtgaat
     4381 ctgttattgt ctcgcctgat gttaagggta ctgtgactgt atattcatct gacgttaaac
     4441 ctgaaaatct acgcaatttc tttatttctg ttttacgtgc aaataatttt gatatggtag
     4501 gttctaaccc ttccattatt cagaagtata atccaaacaa tcaggattat attgatgaat
     4561 tgccatcatc tgataatcag gaatatgatg ataattccgc tccttctggt ggtttctttg
     4621 ttccgcaaaa tgataatgtt actcaaactt ttaaaattaa taacgttcgg gcaaaggatt
     4681 taatacgagt tgtagaattg tttgttaaat ctaatacgtc taaatcctca aatgtattat
     4741 ctattgacgg ctctaatcta ttagttgtta gtgctcctaa agatatttta gataaccttc
     4801 ctcaattcct ttcaactgtt gatttgccaa ctgaccagat attgattgag ggtttgatat
     4861 tcgaggttca gcaaggtgat gctttagatt tttcatttgc tgctggctct cagcgtggca
     4921 ctgttgcagg cggtgttaat actgaccgcc tcacctctgt tttatcttct gctggtggtt
     4981 cgttcggtat ttttaatggc gatgttttag ggctatcagt tcgcgcatta aagactaata
     5041 gccattcaaa aatattgtct gtgccacgta ttcttacgct ttcaggtcag aagggttcta
     5101 tttctgttgg tcagaatgtc ccttttatta ctggtcgtgt gactggtgaa tctgccaatg
     5161 taaataatcc atttcagacg attgagcgtc aaaatgtagg tatttccatg agcgtttttc
     5221 ctgttgcaat ggctggcggt aatattgttc tggacattac cagcaaggcc gatagtttga
     5281 gttcttctac tcaggcaagt gatgttatta ctaatcaaag aagtattgcg acaacggtta
     5341 atttgcgtga tggacagact cttttactcg gtggcctcac tgattataaa aacacttctc
     5401 aggattctgg cgtaccgttc ctgtctaaaa tccctttaat cggcctcctg tttagctccc
     5461 gctctgattc taacgaggaa agcacgttat acgtgctcgt caaagcaacc atagtacgcg
     5521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5641 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5881 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5941 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     6001 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6061 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6121 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6181 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6241 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6301 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6361 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6421 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6481 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6541 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6601 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6661 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6721 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6781 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6841 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6901 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6961 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     7021 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7081 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7141 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7201 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7261 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7321 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7381 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7441 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7501 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gagagaaagg
     7561 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7621 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7681 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7741 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7801 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7861 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     7921 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     7981 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     8041 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     8101 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     8161 gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg
     8221 caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg
     8281 ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct
     8341 gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct
     8401 agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     8461 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     8521 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     8581 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
     8641 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     8701 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     8761 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     8821 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     8881 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     8941 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     9001 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     9061 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     9121 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     9181 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     9241 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     9301 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     9361 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     9421 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     9481 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     9541 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     9601 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     9661 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     9721 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     9781 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     9841 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     9901 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     9961 agtgaacgga tctcgacggt atcgctagct tttaaaagaa aaggggggat tggggggtac
    10021 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
    10081 aaacaaatta caaaaattca aaattttact agtgattatc ggatcaactt tgtatagaaa
    10141 agttgggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
    10201 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg
    10261 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
    10321 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa
    10381 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt
    10441 gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
    10501 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg
    10561 cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg
    10621 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt
    10681 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg
    10741 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
    10801 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg
    10861 tgcctggtct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg
    10921 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat
    10981 ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct
    11041 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
    11101 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg
    11161 cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga
    11221 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc
    11281 agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgacaagtt tgtacaaaaa
    11341 agcaggctgc caccatgttg gtgcgactaa ccaagctgtc ctgccctgcc taccattggt
    11401 ttcatgcctt aaaaattaag aagtgtctac cactatgcgc gccaagatgt tcttcaacct
    11461 ctgctgtaaa aaaaaaaaaa aaaaaacctc agattaccac tcactacact gttcatcccc
    11521 gggagaaaga caaaagatgg gaaggagtga atatggagag gtttgcagaa gaagcagatg
    11581 tggtcatagt tggtgccggc ccagcaggac tctctgcagc tattcggcta aagcagctgg
    11641 ctgctgaaca ggggaaggac atccgtgtgt gtctggtgga gaaagctgct cagatcggag
    11701 cacacacgct ctcaggggct tgtcttgatc cagctgcctt caaagagctc ttcccagact
    11761 ggaaggagaa aggggctcca cttaatactc ctgtaacaga agacaggttt gccattttaa
    11821 cagagaaaca cagaattcct gtgccaattc ttccaggtct tccgatgaac aatcatggca
    11881 attacatcgt acgcctagga catctggtga gctggatggg agaacaggca gaggctcttg
    11941 gagtcgaagt gtatcctggg tatgctgctg ctgaggtcct ttatcatgaa gatggtagtg
    12001 taaaaggaat cgccactaat gatgtaggca tacaaaagga tggtgcacca aagacaacat
    12061 ttgagagagg actggaactg catgccaaag tcacagtctt tgcagagggt tgccatggac
    12121 acctagccaa gcagctttat aagaaatttg atctgagggc cagctgtgat gcacagactt
    12181 acggaattgg tttgaaggag ttatggatta ttgatgaaaa gaagtggaaa cctggaagag
    12241 tagatcacac tgttggttgg cccttggaca gacatactta tggaggctct ttcctttacc
    12301 acttaaatga aggtgaacct ctagtagctg ttggctttgt ggttggcctc gactatcaga
    12361 atccgtacct gagtccattc cgagagttcc agaggtggaa gcatcatccc agcatccagc
    12421 caaccctgga aggcgggaag aggatagcct acggagcccg agcgctcaat gaaggcggct
    12481 tgcagtccat accaaaactc acttttcctg gtggcttact aattggttgt agtcctggtt
    12541 tcatgaatgt ccccaagatc aaaggtaccc acacggcaat gaaaagtgga agtttggcag
    12601 cagaatcaat ttttaagcaa ctaactagtg aaaatctcca gtcaaagaca acaggactcc
    12661 atgtaactga atatgaggac aatttgaagc aatcctgggt atggaaagag ctacacgctg
    12721 ttcgaaatat aaggccatct tgccacggga tactgggagt atatggaggg atgatttaca
    12781 ctgggatatt ttactggata ttgagaggaa tggagccatg gactctaaaa cataaaggct
    12841 cagactctga tcagctcaaa ccagccaagg attgtacacc cattgagtat ccgaaacctg
    12901 atggacagat cagttttgac ctcttatcct ctgtggcttt gagtggtact aatcatgaac
    12961 atgaccagcc agcacattta accttaaagg atgacagcat acctgttaat agaaacctgt
    13021 caatatacga tgggcctgag cagcgattct gccctgcagg agtttatgaa tttgttcctt
    13081 tggaacaagg tgatggattt cgattacaga taaatgctca gaactgtgtg cactgtaaaa
    13141 catgtgatat caaagaccca agtcaaaata ttaactgggt ggtcccagaa ggaggaggag
    13201 ggcctgctta caatggcatg taaacccagc tttcttgtac aaagtggtga taatcgaatt
    13261 ccgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
    13321 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
    13381 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
    13441 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
    13501 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
    13561 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
    13621 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
    13681 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
    13741 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
    13801 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcggg
    13861 aattcccgcg gttcgaattc taccgggtag gggaggcgct tttcccaagg cagtctggag
    13921 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
    13981 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
    14041 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
    14101 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg
    14161 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
    14221 tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg
    14281 gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca
    14341 cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctca cgtggccacc
    14401 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
    14461 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
    14521 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
    14581 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
    14641 ggggaattta gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
    14701 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
    14761 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
    14821 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
    14881 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
    14941 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
    15001 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
    15061 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
    15121 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
    15181 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
    15241 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
    15301 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
    15361 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
    15421 gaatagggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
    15481 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
    15541 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
    15601 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
    15661 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
    15721 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
    15781 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
    15841 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    15901 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
    15961 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
    16021 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
    16081 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
    16141 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
    16201 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
    16261 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
    16321 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
    16381 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
    16441 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
    16501 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
    16561 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
    16621 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggc attttgccaa
    16681 tttcggccta ttggttaaaa aatgagctta tttaacaaaa atttaacgcg aattttaaca
    16741 aaatattaac gtttacaatt taaatatttg cttatacaat cttcctgttt ttggggcttt
    16801 tctgattatc aaccggggta catatgattg acatgctagt tttacgatta ccgttcatcg
    16861 attctcttgt ttgctccaga ctctcaggca atgacctgat agcctttgta gacctctcaa
    16921 aaatagctac cctctccggc atgaatttat cagctagaac ggttgaatat catattgatg
    16981 gtgatttgac tgtctccggc ctttctcacc cgtttgaatc tttacctaca cattactcag
    17041 gcattgcatt taaaatatat gagggttcta aaaattttta tccttgcgtt gaaataaagg
    17101 cttctcccgc aaaagtatta cagggtcata atgtttttgg tacaaccgat ttagctttat
    17161 gctctgaggc tttattgctt aattttgcta attcttt
//
