LOCUS       sequence-492985-       11339 bp    DNA     circular SYN 18-NOV-2025
DEFINITION  Multiplexable FUCCI sensor (FUCCIplex). The EF1-alpha promoter 
            drives the expression of the fluorescent sensor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11339)
  AUTHORS   Di Sante M, Pezzotti M, Zimmermann J, Enrico A, Deschamps J, Balmas 
            E, Becca S, Reali A, Bertero A, Jug F, Pasqualini FS
  TITLE     CALIPERS: Cell cycle-aware live imaging for phenotyping experiments 
            and regeneration studies
  JOURNAL   bioRxiv 2024.12.19.629259
REFERENCE   2  (bases 1 to 11339)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.12.19.629259"
COMMENT     Sequence Label: pLV_[Exp]-Hygro-EF1A-FUCCIplex
FEATURES             Location/Qualifiers
     source          1..11339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(286..306)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_feature    324..441
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(707..751)
                     /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    936..1169
                     /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    1662..1787
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1834..2014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2015..2241)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(2269..2289)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2269..2287)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2308..2324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2308..2324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2321..2343)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2332..2348
                     /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(2356..2362,2363..2380,2381..2386))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2401..2422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2539..2556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2710..3298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2790..2809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3469..4260,4261..4329))
                     /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     4092..4111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4330..4434)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4460..4915)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4597..4618)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4809..4828
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5042..5064
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5056..5073
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5057..5073
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5083..5102
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5083..5101
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      5122..5257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5297..5418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5308..5327)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5362..5381
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             5490..5723
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(5795..6820)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(5795..6820)
                     /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"
     promoter        complement(6837..7336)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6885..6904)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     7278..7301
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    7367..7955
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7473..7484
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7882..7902
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    7973..7997
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(8001..8330)
                     /codon_start=1
                     /product="degron consisting of residues 1-110 of human 
                     Geminin (Zielke and Edgar, 2015)"
                     /label=Gem1 (1-110)
                     /translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS
                     AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE
                     KRRKAL"
     CDS             complement(8361..9305)
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     CDS             complement(9312..9365)
                     /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             complement(9372..9644)
                     /codon_start=1
                     /product="degron consisting of residues 30-120 of human 
                     Cdt1 (Zielke and Edgar, 2015)"
                     /label=Cdt1 (30-120)
                     /translation="PSPARPALRAPASATSGSRKRARPPAAPGRDQARPPARRRLRLSV
                     DEVSSPSTPEAPDIPACPSPGQKIKKSTPAAGQPPHLTSAQDQDTI"
     CDS             complement(join(9699..10409,10410..10412,10413..10415))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9741..9762)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     10090..10109
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10349..10370
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      10412..10421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(10422..10446)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(join(10446..11339,1..285))
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(10455..11339,1..54)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(10478..10498)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 agggccataa cccgtaaaga ggccaggccc gcgggaacca cacacggcac ttacctgtgt
       61 tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac ttatatacgg
      121 ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt cccagtttac
      181 cccgcgccac cttctctagg caccggttca attgccgacc cctcccccca acttctcggg
      241 gactgtgggc gatgtgcgct ctgcccactg acgggcaccg gagcccaact tttctataca
      301 aagttgatcc gataatcact agtaaaattt tgaatttttg taatttgttt ttgtaattct
      361 ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact gtacccccca
      421 atcccccctt ttcttttaaa agctagcgat accgtcgaga tccgttcact aatcgaatgg
      481 atctgtctct gtctctctct ccaccttctt cttctattcc ttcgggcctg tcgggtcccc
      541 tcggggttgg gaggtgggtc tgaaacgata atggtgaata tccctgccta actctattca
      601 ctatagaaag tacagcaaaa actattctta aacctaccaa gcctcctact atcattatga
      661 ataattttat ataccacagc caatttgtta tgttaaacca attccacaaa cttgcccatt
      721 tatctaattc caataattct tgttcattct tttcttgctg gttttgcgat tcttcaatta
      781 aggagtgtat taagcttgtg taattgttaa tttctctgtc ccactccatc caggtcgtgt
      841 gattccaaat ctgttccaga gatttattac tccaactagc attccaaggc acagcagtgg
      901 tgcaaatgag ttttccagag caaccccaaa tccccaggag ctgttgatcc tttaggtatc
      961 tttccacagc caggattctt gcctggagct gcttgatgcc ccagactgtg agttgcaaca
     1021 gatgctgttg cgcctcaata gccctcagca aattgttctg ctgctgcact ataccagaca
     1081 ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg
     1141 ctcccaagaa cccaaggaac aaagctccta ttcccactgc tcttttttct ctctgcacca
     1201 ctcttctctt tgccttggtg ggtgctactc ctaatggttc aatttttact actttatatt
     1261 tatataattc acttctccaa ttgtccctca tatctcctcc tccaggtctg aagatcagcg
     1321 gccgcttgct gtgcggtggt cttacttttg ttttgctctt cctctatctt gtctaaagct
     1381 tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc tactgtatta
     1441 tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg tcccagtatt
     1501 tgtctacagc cttctgatgt ttctaacagg ccaggattaa ctgcgaatcg ttctagctcc
     1561 ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg ccttaaccga
     1621 attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc gcacccatct
     1681 ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg
     1741 cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc ctctggtttc
     1801 cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac actgactaaa
     1861 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
     1921 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
     1981 cccaggctca gatctggtct aaccagagag acccgtttat gtatcgagct aggcacttaa
     2041 atacaatatc tctgcaatgc ggcaattcag tggttcgtcc aatccatgtc agacccgtct
     2101 gttgccttcc taataaggca cgatcgtacc accttacttc caccaatcgg catgcacggt
     2161 gctttttctc tccttgtaag gcatgttgct aactcatcgt taccatgttg caagactaca
     2221 agagtattgc ataagactac attaagcttg cagctccagc ttttgttccc tttagtgagg
     2281 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2341 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2401 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2461 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2521 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2581 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2641 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2701 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2761 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2821 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctctc
     2881 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2941 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3001 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3061 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3121 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3181 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3241 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3301 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3361 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3421 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3481 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3541 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3601 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3661 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3721 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3781 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3841 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3901 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3961 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4021 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4081 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4141 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4201 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4261 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4321 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4381 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4441 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4501 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4561 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4621 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4681 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4741 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4801 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4861 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     4921 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     4981 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     5041 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     5101 ggcgaattgg gtacgtccct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     5161 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     5221 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggatag
     5281 ctagagccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     5341 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     5401 agctgcaata aacaagttcc tctcactctc tgatattcat ttctttgcaa gttataaata
     5461 ctgaataata agatgacatg aactactact gctagagatt ttccacactg actaaaaggg
     5521 tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac
     5581 tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
     5641 ggctcagatc tggtctaacc agagagaccc agtacaagca aaaagcagat cttgtcttcg
     5701 ttgggagtga attagccctt ccagtccccc cttttctttt aaaaagtggc taagatctac
     5761 agctgccttg taagtcattg gtcttaaagg taccctattc ctttgccctc ggacgagtgc
     5821 tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc cagacggccg
     5881 cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg acgattgcgt
     5941 cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag ctctgataga
     6001 gttggtcaag accaatgcgg agcatatacg cccggagccg cggcgatcct gcaagctccg
     6061 gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac ggcctccaga
     6121 agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc cagtcaatga
     6181 ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc gcgtgcacga
     6241 ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga
     6301 cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg ggatcagcaa
     6361 tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt ccgaatgggc
     6421 cgaacccgct cgtctggcta agatcggccg cagcgatcgc atccatggcc tccgcgaccg
     6481 gctgcagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac
     6541 ggcgggagat gcaataggtc aggctctcgc taaattcccc aatgtcaagc acttccggaa
     6601 tcgggagcgc ggccgatgca aagtgccgat aaacataacg atctttgtag aaaccatcgg
     6661 cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg aaagcacgag
     6721 attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa
     6781 acttctcgac agacgtcgcg gtgagttcag gctttttcat ggtggccacg tgaggtcgaa
     6841 aggcccggag atgaggaaga ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag
     6901 aatgccgggc ctccggagga ccttcgggcg cccgccccgc ccctgagccc gcccctgagc
     6961 ccgcccccgg acccacccct tcccagcctc tgagcccaga aagcgaagga gcaaagctgc
     7021 tattggccgc tgccccaaag gcctacccgc ttccattgct cagcggtgct gtccatctgc
     7081 acgagactag tgagacgtgc tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg
     7141 gcggggggga acttcctgac taggggagga gtagaaggtg gcgcgaaggg gccaccaaag
     7201 aacggagccg gttggcgcct accggtggat gtggaatgtg tgcgaggcca gaggccactt
     7261 gtgtagcgcc aagtgcccag cggggctgct aaagcgcatg ctccagactg ccttgggaaa
     7321 agcgcctccc ctacccggta gaattcgaac cgcgggaatt cccgatgcgg ggaggcggcc
     7381 caaagggaga tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc
     7441 ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag
     7501 gacgtcccgc gcagaatcca ggtggcaaca caggcgagca gccatggaaa ggacgtcagc
     7561 ttccccgaca acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg
     7621 gcaaggcagg cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg
     7681 gaaaggagct gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca
     7741 gtgcacacca cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac
     7801 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
     7861 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
     7921 aatctttcac aaattttgta atccagaggt tgattatcgg aattcgatta tcaccacttt
     7981 gtacaagaaa gctgggttta cagggccttc cgccgcttct cggccacctc tttccagtac
     8041 tggctgctgg ggttttcttt gatcatcagg tcgaagcttt cttgggtcac gccgcccagg
     8101 ttcttgttct cgctgctctc gggcacgatc acgcctgggc tagaggtggt gctggtcagg
     8161 tggtcgttcc ggtgcttccg cttgctcagg ccggcagaca gctcattctc ccggcccacc
     8221 agagatccgc tggcgctggg ctggatcatc ttcagggtcc gcctgggcac gctgctgttc
     8281 ttgatgttct ctttgatctc ttcctgcttc tgcttcatgc tggggttcat ttccagaggc
     8341 cgccagtggc tggggtagcc gctctcaagc gcggtgatcc gcgtcgcgat cctttcggcg
     8401 agccgtttgc agatcgcccg cagctcgaaa cggatgaagc gcggcagata atggtgacag
     8461 acaaccaggc cccacagctt gccgccgacc accagcgaca ccgccagggt ggcgcgcacg
     8521 cccatgtcct tcaggaactg cagatggcac ggcgacatcg agcgcaggaa gcagcccgac
     8581 atgtcgagat cgcgcccggt cagcggcgac agccgcggct ccagcggcac cggctgatag
     8641 gtgacgtcga ccagcacgcg gacgcgctgc cgcacgtaca gctgccgcgc catctgcggg
     8701 acagtcgacg acggatagcg gttgccgaaa taggattcga gcccaggcac atggcactcg
     8761 gagaatacca ggccgtggcc ttgctcatcg aaacgataca ccatcacccg gtcgtagccg
     8821 gtgcactgct gaaacagcag cacggtgtca tcgcacagcg cgcgcagtga acccgccgtg
     8881 cggatccgct ccagcgccgg cgccagcgtg cctgacagat cgatcgacgg gccggcacgt
     8941 tcgagttcga tgatcagccc gccttccgga ggccgatgca tcagaccgca gtactccgta
     9001 gagggattgc cgatccggca gcgcaccgcg accggcatgc cttcggcggt gggatcgaga
     9061 tgcggcagga tcttgatcaa cagatcgccg tcgatctcgg cgagcggaac gccgagtacg
     9121 cttccgagat tcagaaattc cgcggcgttg gcgctggcct ggatgacgcg atgatcatgt
     9181 tcgctgacga ccagaagcgc gccatgcggc tggatcgagc cggcgaggtg gatctcttca
     9241 tgttcgcaat tcgaatgaga ggcggtcccg aatgcggggc tgccagaggc atgacctgct
     9301 accatggatc tggggccggg gttctcctcc acgtcgccgc aggtcagcag gctgcccctg
     9361 ccctcgtcga cgatggtgtc ctggtcctgg gcgctggtca ggtgaggagg ctgtccggcg
     9421 gcaggtgtgc tcttcttgat tttctggcca gggctgggac aggcagggat atcgggggcc
     9481 tcgggggtgc tagggctgga cacctcatcc acggacagcc ggagccgtct gcgggcaggg
     9541 ggcctggcct gatctctgcc aggggcggca ggaggtctgg ctctctttct gctgccggag
     9601 gtggcggagg caggggctct cagggcaggt ctggcaggag agggttccag gggcctccaa
     9661 tggctagggt agccgccgag gctaggcagc atgctgtact tgtacagctc gtccatgccg
     9721 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     9781 gggtctttgc tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
     9841 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg
     9901 atgttgtggc ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata
     9961 tagacgttgt cgctaaagta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
    10021 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
    10081 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
    10141 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg
    10201 aagcactgca cgccccagga cagggtggtc acgagggtgg gccagggcac gggcagcttg
    10261 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
    10321 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
    10381 accccggtga acagctcctc gcccttgctc accatggtgg cagcctgctt ttttgtacaa
    10441 acttgtcacg acacctgaaa tggaagaaaa aaactttgaa ccactgtctg aggcttgaga
    10501 atgaaccaag atccaaactc aaaaagggca aattccaagg agaattacat caagtgccaa
    10561 gctggcctaa cttcagtctc cacccactca gtgtggggaa actccatcgc ataaaacccc
    10621 tccccccaac ctaaagacga cgtactccaa aagctcgaga actaatcgag gtgcctggac
    10681 ggcgcccggt actccgtgga gtcacatgaa gcgacggctg aggacggaaa ggcccttttc
    10741 ctttgtgtgg gtgactcacc cgcccgctct cccgagcgcc gcgtcctcca ttttgagctc
    10801 cctgcagcag ggccgggaag cggccatctt tccgctcacg caactggtgc cgaccgggcc
    10861 agccttgccg cccagggcgg ggcgatacac ggcggcgcga gaccaggcac cagagcaggc
    10921 cggccagctt gagactaccc ccgtccgatt ctcggtggcc gcgctcgcag gccccgcctc
    10981 gccgaacatg tgcgctggga cgcacgggcc ccgtcgccgc ccgcggcccc aaaaaccgaa
    11041 ataccagtgt gcagatcttg gcccgcattt acaagactat cttgccagaa aaaaagcgtc
    11101 gcagcaggtc atcaaaaatt ttaaatggct agagacttat cgaaagcagc gagacaggcg
    11161 cgaaggtgcc accagattcg cacgcggcgg ccccagcgcc caggccaggc ctcaactcaa
    11221 gcacgaggcg aaggggctcc ttaagcgcaa ggcctcgaac tctcccaccc acttccaacc
    11281 cgaagctcgg gatcaagaat cacgtactgc agccaggtgg aagtaattca aggcacgca
//
