LOCUS       sequence-495380-        8890 bp    DNA     circular SYN 16-DEC-2025
DEFINITION  A PiggyBac expression vector containing the human EF1a promoter 
            driving FastFUCCI to label cell cycle state followed by an 
            IRES-3xnls-mTagBFP2. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8890)
  AUTHORS   Gillespie WB 3rd, Zhang Y, Ruiz OE, Cerda J 3rd, Ortiz-Guzman J, 
            Sherman M, Turner WD, Largoza G, Mosser LE, Fujimoto E, Chien CB, 
            Kwan KM, Arenkiel BR, Devine WP, Wythe JD
  TITLE     MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox for use in vertebrate model systems.
  JOURNAL   Development. 2025 Nov 15;152(22):dev204308. doi: 10.1242/dev.204308.
            Epub 2025 Nov 13.
  PUBMED    41230856
REFERENCE   2  (bases 1 to 8890)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Development."; date: "2025-11-15"; volume: "152(22):dev204308. 
            doi"; pages: " 10.1242/dev.204308. Epub 2025 Nov 13"
COMMENT     Sequence Label: pB-EF1a-FastFUCCI-IRES-3xnls-mTagBFP2 (JDW 1511)
FEATURES             Location/Qualifiers
     source          1..8890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   660..694
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1002..1022
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1024..1042
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1067..1083
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1068..1084
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1173..2351
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1404..2342
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2299..2319
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(2359..2375)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2359..2375)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2412..2430)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2430..2454
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2496..3146
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     CDS             3201..3473
                     /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             3480..3533
                     /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             3540..4217
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     CDS             3540..4217
                     /codon_start=1
                     /product="monomeric Azami-Green fluorescent protein 
                     (Karasawa et al., 2003)"
                     /label=mAzami-Green
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     CDS             4248..4577
                     /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"
     protein_bind    complement(4607..4631)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(4632..5206,5207..5209,5210..5218)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4799..4816)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5026..5045
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5222..5287
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5306..5326
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5330..5350
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5354..5374
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5396..6106
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    6163..6297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6187..6206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6241..6260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(6359..6379)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(6387..6403)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   6616..6678
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(6702..6720)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6741..6757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6741..6757)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6754..6776)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6765..6781
                     /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(6789..6795,6796..6813,6814..6819))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6834..6855
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6972..6989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7143..7731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7223..7242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7902..8693,8694..8762))
                     /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     8525..8544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8763..8867)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccttaac
      661 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
      721 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
      781 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg
      841 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat
      901 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta
      961 tagatatcgt cgacttaatt aacatgatta cgccaagcta tcaactttgt atagaaaagt
     1021 tggtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga ggtcgacggt
     1081 atcgataagc tttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat
     1141 ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc
     1201 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
     1261 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     1321 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     1381 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct
     1441 ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc
     1501 ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag
     1561 ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa
     1621 tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt
     1681 tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag
     1741 atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc
     1801 agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt
     1861 agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc
     1921 cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc
     1981 ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg
     2041 agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca
     2101 cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt
     2161 ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga
     2221 ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt
     2281 ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc
     2341 aggtgtcgtg aggtaccgga tccactagtt ctagagcggc cgccaccgcg gtggagctcc
     2401 agcttttgtt ccctttagtg agggttaatt caagtttgta caaaaaagca ggctggtctg
     2461 actggtacct cgtgattaat taagaattgg gatccatggt gtccgtgatc aagcccgaga
     2521 tgaagatgcg gtactacatg gacggcagcg tgaacggcca cgagttcacc atcgagggcg
     2581 agggcaccgg cagaccttac gagggccacc aggaaatgac cctgagagtc accatggccg
     2641 agggcggacc catgcccttc gccttcgacc tggtgtccca cgtgttctgc tacggccacc
     2701 gggtgttcac caagtacccc gaggaaatcc ccgactactt caagcaggcc ttccccgagg
     2761 gcctgagctg ggagcggagc ctggaattcg aggatggcgg cagcgcctct gtgtctgccc
     2821 acatcagcct gcggggcaac accttctacc acaagagcaa gttcaccggc gtgaacttcc
     2881 ccgccgacgg ccccatcatg cagaaccaga gcgtggactg ggagcccagc accgagaaga
     2941 tcaccgccag cgacggcgtg ctgaagggcg acgtgaccat gtacctgaag ctggaaggcg
     3001 gcggaaacca caagtgccag atgaaaacca cctacaaggc cgccaaagaa atcctggaaa
     3061 tgcccggcga ccactacatc ggccaccggc tggtgcggaa aaccgagggc aacatcaccg
     3121 agcaggtgga ggatgccgtg gcccactaca gcatgctgcc tagcctcggc ggctacccta
     3181 gccattggag gcccctggaa ccctctcctg ccagacctgc cctgagagcc cctgcctccg
     3241 ccacctccgg cagcagaaag agagccagac ctcctgccgc ccctggcaga gatcaggcca
     3301 ggccccctgc ccgcagacgg ctccggctgt ccgtggatga ggtgtccagc cctagcaccc
     3361 ccgaggcccc cgatatccct gcctgtccca gccctggcca gaaaatcaag aagagcacac
     3421 ctgccgccgg acagcctcct cacctgacca gcgcccagga ccaggacacc atcgtcgacg
     3481 agggcagggg cagcctgctg acctgcggcg acgtggagga gaaccccggc cccagatcca
     3541 tggtgtccgt gatcaagccc gagatgaaga tcaagctgtg catgcgggga accgtgaacg
     3601 gccacaactt cgtgatcgag ggcgagggca agggaaaccc ctacgagggc acccagatcc
     3661 tggacctgaa cgtgaccgaa ggcgcccctc tgcccttcgc ctacgacatc ctgaccaccg
     3721 tgttccagta cggcaaccgg gccttcacca agtaccccgc cgacatccag gactacttca
     3781 agcagacctt ccccgagggc taccactggg agcggagcat gacctacgag gaccagggca
     3841 tctgcaccgc cacctccaac atcagcatga ggggcgactg ctttttctac gacatcagat
     3901 tcgacggcac caacttcccc cccaacggcc ccgtgatgca gaaaaagacc ctgaagtggg
     3961 agcccagcac cgagaagatg tacgtggagg acggcgtgct gaagggcgac gtgaacatgc
     4021 ggctgctgct ggaaggcggc ggacactaca gatgcgactt caagaccacc tacaaggcca
     4081 agaaagaagt gcggctgccc gacgcccaca agatcgacca ccggatcgag atcctgaagc
     4141 acgacaagga ctacaacaaa gtgaagctgt acgagaacgc cgtggccagg tacagcatgc
     4201 tgcccagcca ggccaagggc taccccagcc actggcggcc tctggaaatg aaccccagca
     4261 tgaagcagaa gcaggaagag atcaaagaga acatcaagaa cagcagcgtg cccaggcgga
     4321 ccctgaagat gatccagccc agcgccagcg gatctctggt gggccgggag aatgagctgt
     4381 ctgccggcct gagcaagcgg aagcaccgga acgaccacct gaccagcacc acctctagcc
     4441 caggcgtgat cgtgcccgag agcagcgaga acaagaacct gggcggcgtg acccaagaaa
     4501 gcttcgacct gatgatcaaa gaaaacccca gcagccagta ctggaaagag gtggccgaga
     4561 agcggcggaa ggccctgtag agatctgact gagcggccgc cagtcaaccc agctttcttg
     4621 tacaaagtgg ccccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     4681 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     4741 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     4801 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     4861 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     4921 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     4981 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     5041 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     5101 gggcctcggt acacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     5161 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     5221 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     5281 tgacaaagga tccaccggtg aattcccgaa gaagaaaaga aaagtaggac ccaagaagaa
     5341 acgcaaagtc ggaccgaaga agaaacgaaa agtcagatct ggcggtgctg gtggtatggt
     5401 gtctaagggc gaagagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac
     5461 cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
     5521 ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct
     5581 ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg gcatccccga
     5641 cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga
     5701 cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa
     5761 cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga agaaaacact
     5821 cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg aaggcagaaa
     5881 cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacgcca agaccacata
     5941 tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg tggactacag
     6001 actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg aggtggcagt
     6061 ggccagatac tgcgacctcc ctagcaaact ggggcacaag ctcaatgcta gcggctgata
     6121 tctataatga ggcgcgccgt cgacacgcgt taatgatacg tagatccaga catgataaga
     6181 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     6241 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     6301 aacatctaga cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttcca
     6361 actttattat acatagttga taattcactg gccgtcgttt tacggtacca tcgatgatga
     6421 taagcttgtt taaacgttaa cggatccttt gttactttat agaagaaatt ttgagttttt
     6481 gttttttttt aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat
     6541 gtaagtgtaa atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata
     6601 cagaccgata aaacacatgc gtcaatttta cgcatgatta tctttaacgt acgtcacaat
     6661 atgattatct ttctagggtt aaagagctcc agcttttgtt ccctttagtg agggttaatt
     6721 tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     6781 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6841 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6901 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6961 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7021 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7081 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7561 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7621 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7681 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7741 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7801 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7861 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7921 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7981 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     8041 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8101 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8161 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8221 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8281 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8341 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8401 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8461 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8521 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8581 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8641 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8701 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8761 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8821 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8881 aaagtgccac
//
