LOCUS       sequence-413226-       14581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LV REPORT containing minimal promoter (GtwyA inserted into MCS) 
            driving monomeric nuclear Tomato followed by an PKG/CMV promoter 
            driving nuclear d2eGFP E2A Hygromycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14581)
  AUTHORS   Allen TP, Roennfeldt AE, Reckdharajkumar M, Liu M, Quinn RJ, Russell
            DL, Peet DJ, Whitelaw ML, Bersten DC
  TITLE     dFLASH; dual FLuorescent transcription factor Activity Sensor for 
            Histone integrated live-cell reporting and high-content screening
  JOURNAL   bioRxiv 2023.11.21.568191
REFERENCE   2  (bases 1 to 14581)
  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: "bioRxiv 
            2023.11.21.568191"
COMMENT     Sequence Label: pLV-REPORT(PGK/CMV)-GtwyA
FEATURES             Location/Qualifiers
     source          1..14581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    159..180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(195..200,201..218,219..225)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    233..249
                     /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     238..260
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     257..273
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     257..273
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     292..312
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        294..312
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        340..566
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             567..747
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    794..919
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1412..1645
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1830..1874
                     /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    2136..2253
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2280..2371
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2320..2339
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    2391..2515
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2540..2545,2546..2563,2564..2570)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2624..3283
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2690..2709)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3625..3930
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3809..3828
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3971..4095)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     4162..4186
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4291..4992
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(4424..4442)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4692..4711
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4886..4909
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             5002..5022
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5026..5046
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5050..5070
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5119..6246
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     CDS             6247..6300
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6295..7095
                     /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="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6355..6374)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6965..6984
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7140..7163)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7490..7509
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(7612..7631)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7803..8182
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8183..8386
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8337..8357
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8431..8450
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8431..8449
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    8479..8503
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      8530..8539
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8551..8571
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8575..8595
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8599..8619
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(8629..8631,8632..8634,8635..9345,9346..9351,
                     9352..9471)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(8674..8695)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8935..8954)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             9472..9531
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             9532..10557
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             9532..10557
                     /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             11790..12023
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    12095..12216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12132..12151)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12186..12205
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     12318..12337
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(12602..12620)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        12688..12792
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12793..12861,12862..13653)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(13011..13030)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      13824..14412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14313..14332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgataaccct aattcgatag catatgcttc ccgttgggta acatatgcta ttgaattagg
       61 gttagtctgg atagtatata ctactacccg ggaagcatat gctacccgtt tagggttcac
      121 cggtgatgcc ggccacgatg cgtccggcgt agaggatcta atgtgagtta gctcactcat
      181 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      241 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      301 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
      361 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
      421 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
      481 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
      541 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
      601 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
      661 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
      721 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
      781 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
      841 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
      901 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
      961 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     1021 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     1081 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     1141 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     1201 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     1261 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     1321 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     1381 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     1441 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     1501 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     1561 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     1621 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     1681 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     1741 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     1801 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     1861 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     1921 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     1981 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2041 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     2101 ccattcgatt agtgaacgga tctcgacggt atcggtttta aaagaaaagg ggggattggg
     2161 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     2221 ttacaaaaac aaattacaaa aattcaaaat tttatcgata tctagaactc gagagtacta
     2281 acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc
     2341 agtgcaagtg caggtgccag aacatttctc tggcctaact ggccggtacc acaagtttgt
     2401 acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt
     2461 tgcataaaaa acagactaca taatactgtg aaacacaaca tatccagtca ctatggcggc
     2521 cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg tgtggatttt
     2581 gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     2641 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     2701 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     2761 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     2821 gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga
     2881 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
     2941 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     3001 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc
     3061 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     3121 cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     3181 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
     3241 tgaattacaa cagtactgcg atgagtggca gggcggggcg taaacgcgtg gatccggctt
     3301 actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata
     3361 tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca
     3421 gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg
     3481 gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag
     3541 cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg
     3601 ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc
     3661 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg
     3721 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg
     3781 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
     3841 gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
     3901 gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct
     3961 gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt
     4021 atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt
     4081 cttgtacaaa gtggtagatc tccctagggt ttaaacggcg cgccagcttt aggcgtgtac
     4141 ggtgggcgcc tataaaagca gagctcgttt agtgaaccgt cagatcgcct ggagcaattc
     4201 cacaacactt ttgtcttata ccaactttcc gtaccacttc ctaccctcgt aaagtcgaca
     4261 ccggggccca gatctatcga tcggccggat atggtgagca agggcgagga ggtcatcaaa
     4321 gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc
     4381 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     4441 aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc
     4501 aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag
     4561 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag
     4621 gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc
     4681 ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc
     4741 ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac
     4801 ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg
     4861 cccggctact actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc
     4921 atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta cggcatggac
     4981 gagctgtaca aggccgtgga tccaaaaaag aagagaaagg tagatccaaa aaagaagaga
     5041 aaggtagatc caaaaaagaa gagaaaggta cacggcgcca cgaagcaagc aggagatgtt
     5101 gaagaaaacc ccgggcctat ggcttcgtac cccggccatc aacacgcgtc tgcgttcgac
     5161 caggctgcgc gttctcgcgg ccatagcaac cgacgtacgg cgttgcgccc tcgccggcag
     5221 caagaagcca cggaagtccg cccggagcag aaaatgccca cgctactgcg ggtttatata
     5281 gacggtcccc acgggatggg gaaaaccacc accacgcaac tgctggtggc cctgggttcg
     5341 cgcgacgata tcgtctacgt acccgagccg atgacttact ggcgggtgct gggggcttcc
     5401 gagacaatcg cgaacatcta caccacacaa caccgcctcg accagggtga gatatcggcc
     5461 ggggacgcgg cggtggtaat gacaagcgcc cagataacaa tgggcatgcc ttatgccgtg
     5521 accgacgccg ttctggctcc tcatatcggg ggggaggctg ggagctcaca tgccccgccc
     5581 ccggccctca ccctcatctt cgaccgccat cccatcgccg ccctcctgtg ctacccggcc
     5641 gcgcggtacc ttatgggcag catgaccccc caggccgtgc tggcgttcgt ggccctcatc
     5701 ccgccgacct tgcccggcac caacatcgtg cttggggccc ttccggagga cagacacatc
     5761 gaccgcctgg ccaaacgcca gcgccccggc gagcggctgg acctggctat gctggctgcg
     5821 attcgccgcg tttacgggct acttgccaat acggtgcggt atctgcagtg cggcgggtcg
     5881 tggcgggagg actggggaca gctttcgggg acggccgtgc cgccccaggg tgccgagccc
     5941 cagagcaacg cgggcccacg accccatatc ggggacacgt tatttaccct gtttcgggcc
     6001 cccgagttgc tggcccccaa cggcgacctg tataacgtgt ttgcctgggc cttggacgtc
     6061 ttggccaaac gcctccgttc catgcacgtc tttatcctgg attacgacca atcgcccgcc
     6121 ggctgccggg acgccctgct gcaacttacc tccgggatgg tccagaccca cgtcaccacc
     6181 cccggctcca taccgacgat atgcgacctg gcgcgcacgt ttgcccggga gatgggggag
     6241 gctaacgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct
     6301 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     6361 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     6421 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6481 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6541 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6601 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6661 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6721 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6781 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6841 ggcgatgatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6901 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6961 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     7021 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     7081 gacgagttct tctgaaattc taccgggtag gggaggcgct tttcccaagg cagtctggag
     7141 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     7201 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     7261 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
     7321 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg
     7381 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
     7441 tctgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
     7501 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc
     7561 tgcatctcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     7621 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     7681 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     7741 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     7801 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     7861 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     7921 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     7981 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     8041 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     8101 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     8161 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     8221 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     8281 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     8341 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag
     8401 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
     8461 tggctagtta agctatcaac aagtttgtac aaaaaagcag gctttaaagg aaccaattca
     8521 gtcgaggtcg ccaccatggt gcacgtggat ccaaaaaaga agagaaaggt agatccaaaa
     8581 aagaagagaa aggtagatcc aaaaaagaag agaaaggtac acgtgtccat ggtgagcaag
     8641 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     8701 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     8761 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     8821 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     8881 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     8941 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     9001 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     9061 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     9121 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     9181 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     9241 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     9301 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagaagct tagccatggc
     9361 ttcccgccgg aggtggagga gcaggatgat ggcacgctgc ccatgtcttg tgcccaggag
     9421 agcgggatgg accgtcaccc tgcagcctgt gcttctgcta ggatcaatgt gcagtgtact
     9481 aattatgctc tcttgaaatt ggctggagat gttgagagca acccaggtcc catgaaaaag
     9541 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
     9601 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
     9661 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
     9721 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc
     9781 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
     9841 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct
     9901 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
     9961 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
    10021 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
    10081 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
    10141 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
    10201 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
    10261 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
    10321 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
    10381 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
    10441 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
    10501 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaataggct
    10561 agcctagtgc catttgttca gtggttcgta gggctttccc ccactgtttg gctttcagtt
    10621 atatggatga tgtggtattg ggggccaagt ctgtacagca tcttgagtcc ctttttaccg
    10681 ctgttaccaa ttttcttttg tctttgggta tacatttaaa ccctaacaaa acaaagagat
    10741 ggggttactc tctaaatttt atgggttatg tcattggatg ttatgggtcc ttgccacaag
    10801 aacacatcat acaaaaaatc aaagaatgtt ttagaaaact tcctattaac aggcctattg
    10861 attggaaagt atgtcaacga attgtgggtc ttttgggttt tgctgcccct tttacacaat
    10921 gtggttatcc tgcgttgatg cctttgtatg catgtattca atctaagcag gctttcactt
    10981 tctcgccaac ttacaaggcc tttctgtgta aacaatacct gaacctttac cccgttgccc
    11041 ggcaacggcc aggtctgtgc caagtgtttg ctgacgcaac ccccactggc tggggcttgg
    11101 tcatgggcca tcagcgcatg cgtggaacct tttcggctcc tctgccgatc catactgcgg
    11161 aactcctagc cgcttgtttt gctcgcagca ggtctggagc aaacattatc gggactgata
    11221 actctgttgt cctatcccgc aaatatacat cgtttccatg gctgctaggc tgtgctgcca
    11281 actggatcct gcgcgggacg tcctttgttt acgtcccgtc ggcgctgaat cctgcggacg
    11341 acccttctcg gggtcgcttg ggactctctc gtccccttct ccgtctgccg ttccgaccga
    11401 ccacggggcg cacctctctt tacgcggact ccccgtctgt gccttctcat ctgccggacc
    11461 gtgtgcactt cgcttcacct ctgcacgtcg catggagacc accgtgaacg cccaccaaat
    11521 attgcccaag gtcttacata agaggactct tggactctca gcaatgtcaa cgaccgacct
    11581 tgaggcatac ttcaaagact gtttgtttaa agactgggag gagttggggg aggagattag
    11641 gttaaaggtc tttgtactag gaggctgtag gcataaattg gtctgcgcac cagcaccatg
    11701 gcgcaatcac tagagcgggg tacctttaag accaatgact tacaaggcag ctgtagatct
    11761 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
    11821 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
    11881 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
    11941 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
    12001 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
    12061 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
    12121 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
    12181 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
    12241 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
    12301 ggctgactaa ttttttttat ttatgcagag gccgaggccg gatcccttga gtggctttca
    12361 tcctggagca gactttgcag tctgtggact gcaacacaac attgccttta tgtgtaactc
    12421 ttggctgaag ctcttacacc aatgctgggg gacatgtacc tcccaggggc ccaggaagac
    12481 tacgggaggc tacaccaacg tcaatcagag gggcctgtgt agctaccgat aagcggaccc
    12541 tcaagagggc attagcaata gtgtttataa ggcccccttg ttaattcttg aagacgaaag
    12601 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
    12661 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
    12721 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
    12781 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
    12841 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
    12901 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
    12961 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
    13021 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
    13081 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
    13141 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
    13201 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
    13261 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
    13321 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
    13381 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
    13441 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
    13501 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
    13561 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
    13621 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
    13681 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
    13741 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
    13801 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
    13861 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
    13921 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
    13981 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
    14041 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
    14101 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
    14161 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
    14221 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
    14281 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
    14341 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
    14401 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
    14461 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
    14521 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
    14581 g
//
