LOCUS       sequence-354701-       10740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LV REPORT containing minimal promoter driving monomeric nuclear 
            Tomato followed by an EF1a promoter driving nuclear d2eGFP E2A 
            Hygromycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10740)
  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 10740)
  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(EF1a)
FEATURES             Location/Qualifiers
     source          1..10740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..197
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     437..458
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(513..533)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(537..557)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(561..581)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      593..602
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(681..1867)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          690..1636
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(713..733)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             complement(2058..2078)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2082..2102)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2106..2126)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2136..2837)
                     /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(2219..2242)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2417..2436)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2686..2704
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2942..2966)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    3038..3155
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(3417..3461)
                     /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    3646..3879
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    4372..4497
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4544..4724
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(4725..4951)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(4979..4999)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4979..4997)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5018..5034)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5018..5034)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5031..5053)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5042..5058
                     /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(5066..5072,5073..5090,5091..5096))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5111..5132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5460..6048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5540..5559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6219..7010,7011..7079))
                     /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     6842..6861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7080..7184)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7252..7270
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7535..7554)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    7656..7777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7667..7686)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7721..7740
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             7849..8082
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(9315..10340)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(9315..10340)
                     /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"
     CDS             complement(10341..10400)
                     /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             complement(join(10401..10520,10521..10526,10527..10740,
                     1..497,498..500,501..503))
                     /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(10569..10590)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
       61 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      121 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      181 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
      241 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
      301 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
      361 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
      421 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
      481 agctcctcgc ccttgctcac catggacacg tgtacctttc tcttcttttt tggatctacc
      541 tttctcttct tttttggatc tacctttctc ttcttttttg gatccacgtg caccatggtg
      601 gcgacctcga gaattccagc acactggcgg ccgttactag tggatccgag ctcggtacca
      661 agcttggtca agctaattcc tcacgacacc tgaaatggaa gaaaaaaact ttgaaccact
      721 gtctgaggct tgagaatgaa ccaagatcca aactcaaaaa gggcaaattc caaggagaat
      781 tacatcaagt gccaagctgg cctaacttca gtctccaccc actcagtgtg gggaaactcc
      841 atcgcataaa acccctcccc ccaacctaaa gacgacgtac tccaaaagct cgatcgagaa
      901 ctaatcgagg tgcctggacg gcgcccggta ctccgtggag tcacatgaag cgacggctga
      961 ggacggaaag gcccttttcc tttgtgtggg tgactcaccc gcccgctctc ccgagcgccg
     1021 cgtcctccat tttgagctcc ctgcagcagg gccgggaagc ggccatcttt ccgctcacgc
     1081 aactggtgcc gaccgggcca gccttgccgc ccagggcggg gcgatacacg gcggcgcgag
     1141 gccaggcacc agagcaggcc ggccagcttg agactacccc cgtccgattc tcggtggccg
     1201 cgctcgcagg ccccgcctcg ccgaacatgt gcgctgggac gcacgggccc cgtcgccgcc
     1261 cgcggcccca aaaaccgaaa taccagtgtg cagatcgatc ttggcccgca tttacaagac
     1321 tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg gctagagact
     1381 tatcgaaagc agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc
     1441 gcccaggcca ggcctcaact caagcacgag gcgaaggggc tccttaagcg caaggcctcg
     1501 aactctccca cccacttcca acccgaagct cgggatcaag aatcacgtac tgcagccagg
     1561 tggaagtaat tcaaggcacg caagggccat aacccgtaaa gaggccaggc ccgcgggaac
     1621 cacacacggc acttacctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg
     1681 cgactactgc acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca
     1741 cgacatcact ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga
     1801 cccctccccc caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac
     1861 cggagccaat tcccactcct ttcaagacct agaaggtcca ttagctgcaa agattcctct
     1921 ctgtttaaaa ctttatccat ctttgcaaag ctcggtcgag atctcccgat ccgtcgaggt
     1981 cgacggtcgc tcctttcggt cgggcgctgc gggtcgtggg gcgggcgctg cgggtcgtgg
     2041 ggcgggcgtc agccgtgtac ctttctcttc ttttttggat ctacctttct cttctttttt
     2101 ggatctacct ttctcttctt ttttggatcc acggccttgt acagctcgtc catgccgtac
     2161 aggaacaggt ggtggcggcc ctcggagcgc tcgtactgtt ccacgatggt gtagtcctcg
     2221 ttgtgggagg tgatgtccag cttggtgtcc acgtagtagt agccgggcag ttgcacgggc
     2281 ttcttggcca tgtagatggt cttgaactcc accaggtagt ggccgccgtc cttcagcttc
     2341 agggcctggt ggatctcgcc cttcagcacg ccgtcgcggg ggtacaggcg ctcggtggag
     2401 gcctcccagc ccatggtctt cttctgcatt acggggccgt cgggggggaa gttggtgccg
     2461 cgcatcttca ccttgtagat cagcgtgccg tcctgcaggg aggagtcctg ggtcacggtc
     2521 accagaccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac
     2581 agcttcttgt aatcggggat gtcggcgggg tgcttcacgt acgccttgga gccgtacatg
     2641 aactgggggg acaggatgtc ccaggcgaag ggcagggggc cgcccttggt caccttcagc
     2701 ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg
     2761 tggccgttca tggagccctc catgcgcacc ttgaagcgca tgaactcttt gatgacctcc
     2821 tcgcccttgc tcaccatatc cggccgatcg atagatctgg gccccggtgt cgactttacg
     2881 agggtaggaa gtggtacgga aagttggtat aagacaaaag tgttgtggaa ttgctccagg
     2941 cgatctgacg gttcactaaa cgagctctgc ttttataggc gcccaccgta cacgcctaaa
     3001 gctggcgcgc cgtttaaacc ctaggttcga aatcgataaa attttgaatt tttgtaattt
     3061 gtttttgtaa ttctttagtt tgtatgtctg ttgctattat gtctactatt ctttcccctg
     3121 cactgtaccc cccaatcccc ccttttcttt taaaaccgat accgtcgaga tccgttcact
     3181 aatcgaatgg atctgtctct gtctctctct ccaccttctt cttctattcc ttcgggcctg
     3241 tcgggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata tccctgccta
     3301 actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa gcctcctact
     3361 atcattatga ataattttat ataccacagc caatttgtta tgttaaacca attccacaaa
     3421 cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg gttttgcgat
     3481 tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc ccactccatc
     3541 caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc attccaaggc
     3601 acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag ctgttgatcc
     3661 tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc ccagactgtg
     3721 agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg ctgctgcact
     3781 ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg
     3841 cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc tcttttttct
     3901 ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc aatttttact
     3961 actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc tccaggtctg
     4021 aagatcagcg gccgcttgct gtgcggtggt cttacttttg ttttgctctt cctctatctt
     4081 gtctaaagct tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc
     4141 tactgtatta tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg
     4201 tcccagtatt tgtctacagc cttctgatgt ttctaacagg ccaggattaa ctgcgaatcg
     4261 ttctagctcc ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg
     4321 ccttaaccga attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc
     4381 gcacccatct ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg
     4441 ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc
     4501 ctctggtttc cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac
     4561 actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac
     4621 actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag
     4681 ccagagagct cccaggctca gatctggtct aaccagagag acccgtttat gtatcgagct
     4741 aggcacttaa atacaatatc tctgcaatgc ggcaattcag tggttcgtcc aatccatgtc
     4801 agacccgtct gttgccttcc taataaggca cgatcgtacc accttacttc caccaatcgg
     4861 catgcacggt gctttttctc tccttgtaag gcatgttgct aactcatcgt taccatgttg
     4921 caagactaca agagtattgc ataagactac attaagcttg cagctccagc ttttgttccc
     4981 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5041 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5101 ggggtgccta atgagtgagc taactcacat tagatcctct acgccggacg catcgtggcc
     5161 ggcatcaccg gtgaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat
     5221 ccagactaac cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt
     5281 agggttatcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     5341 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     5401 ggacagcttc aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5461 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5521 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5581 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5641 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5701 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5761 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5821 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5881 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5941 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6361 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     6541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     6841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7201 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     7261 atcacgaggc cctttcgtct tcaagaatta acaagggggc cttataaaca ctattgctaa
     7321 tgccctcttg agggtccgct tatcggtagc tacacaggcc cctctgattg acgttggtgt
     7381 agcctcccgt agtcttcctg ggcccctggg aggtacatgt cccccagcat tggtgtaaga
     7441 gcttcagcca agagttacac ataaaggcaa tgttgtgttg cagtccacag actgcaaagt
     7501 ctgctccagg atgaaagcca ctcaagggat ccggcctcgg cctctgcata aataaaaaaa
     7561 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     7621 ggagttaggg gcgggatagc tagagccaga catgataaga tacattgatg agtttggaca
     7681 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     7741 tttatttgta accattataa gctgcaataa acaagttcct ctcactctct gatattcatt
     7801 tctttgcaag ttataaatac tgaataataa gatgacatga actactactg ctagagattt
     7861 tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg
     7921 cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta
     7981 gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca gtacaagcaa
     8041 aaagcagatc ttgtcttcgt tgggagtgaa ttagcccttc cagtcccccc ttttctttta
     8101 aaaagtggct aagatctaca gctgccttgt aagtcattgg tcttaaaggt accccgctct
     8161 agtgattgcg ccatggtgct ggtgcgcaga ccaatttatg cctacagcct cctagtacaa
     8221 agacctttaa cctaatctcc tcccccaact cctcccagtc tttaaacaaa cagtctttga
     8281 agtatgcctc aaggtcggtc gttgacattg ctgagagtcc aagagtcctc ttatgtaaga
     8341 ccttgggcaa tatttggtgg gcgttcacgg tggtctccat gcgacgtgca gaggtgaagc
     8401 gaagtgcaca cggtccggca gatgagaagg cacagacggg gagtccgcgt aaagagaggt
     8461 gcgccccgtg gtcggtcgga acggcagacg gagaagggga cgagagagtc ccaagcgacc
     8521 ccgagaaggg tcgtccgcag gattcagcgc cgacgggacg taaacaaagg acgtcccgcg
     8581 caggatccag ttggcagcac agcctagcag ccatggaaac gatgtatatt tgcgggatag
     8641 gacaacagag ttatcagtcc cgataatgtt tgctccagac ctgctgcgag caaaacaagc
     8701 ggctaggagt tccgcagtat ggatcggcag aggagccgaa aaggttccac gcatgcgctg
     8761 atggcccatg accaagcccc agccagtggg ggttgcgtca gcaaacactt ggcacagacc
     8821 tggccgttgc cgggcaacgg ggtaaaggtt caggtattgt ttacacagaa aggccttgta
     8881 agttggcgag aaagtgaaag cctgcttaga ttgaatacat gcatacaaag gcatcaacgc
     8941 aggataacca cattgtgtaa aaggggcagc aaaacccaaa agacccacaa ttcgttgaca
     9001 tactttccaa tcaataggcc tgttaatagg aagttttcta aaacattctt tgattttttg
     9061 tatgatgtgt tcttgtggca aggacccata acatccaatg acataaccca taaaatttag
     9121 agagtaaccc catctctttg ttttgttagg gtttaaatgt atacccaaag acaaaagaaa
     9181 attggtaaca gcggtaaaaa gggactcaag atgctgtaca gacttggccc ccaataccac
     9241 atcatccata taactgaaag ccaaacagtg ggggaaagcc ctacgaacca ctgaacaaat
     9301 ggcactaggc tagcctattc ctttgccctc ggacgagtgc tggggcgtcg gtttccacta
     9361 tcggcgagta cttctacaca gccatcggtc cagacggccg cgcttctgcg ggcgatttgt
     9421 gtacgcccga cagtcccggc tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa
     9481 gctgcatcat cgaaattgcc gtcaaccaag ctctgataga gttggtcaag accaatgcgg
     9541 agcatatacg cccggagccg cggcgatcct gcaagctccg gatgcctccg ctcgaagtag
     9601 cgcgtctgct gctccataca agccaaccac ggcctccaga agaagatgtt ggcgacctcg
     9661 tattgggaat ccccgaacat cgcctcgctc cagtcaatga ccgctgttat gcggccattg
     9721 tccgtcagga cattgttgga gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag
     9781 tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg
     9841 tccatcacag tttgccagtg atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc
     9901 catgtagtgt attgaccgat tccttgcggt ccgaatgggc cgaacccgct cgtctggcta
     9961 agatcggccg cagcgatcgc atccatggcc tccgcgaccg gctgcagaac agcgggcagt
    10021 tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac ggcgggagat gcaataggtc
    10081 aggctctcgc tgaattcccc aatgtcaagc acttccggaa tcgggagcgc ggccgatgca
    10141 aagtgccgat aaacataacg atctttgtag aaaccatcgg cgcagctatt tacccgcagg
    10201 acatatccac gccctcctac atcgaagctg aaagcacgag attcttcgcc ctccgagagc
    10261 tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg
    10321 gtgagttcag gctttttcat gggacctggg ttgctctcaa catctccagc caatttcaag
    10381 agagcataat tagtacactg cacattgatc ctagcagaag cacaggctgc agggtgacgg
    10441 tccatcccgc tctcctgggc acaagacatg ggcagcgtgc catcatcctg ctcctccacc
    10501 tccggcggga agccatggct aagcttcttg tacagctcgt ccatgccgag agtgatcccg
    10561 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
    10621 agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
    10681 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
//
