LOCUS       sequence-473566-       11481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A piggybac-based cloning vector containing TRE3G promoter-driven 
            short-form of TP53 and CAG promoter-driven nuclear-localized 
            Clover-P2A-rtTA-IRES-BSD..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11481)
  AUTHORS   Okubo C, Nakamura M, Sato M, Shichino Y, Mito M, Takashima Y, 
            Iwasaki S, Takahashi K
  TITLE     EIF3D safeguards the homeostasis of key signaling pathways in human 
            primed pluripotency.
  JOURNAL   Sci Adv. 2025 Apr 11;11(15):eadq5484. doi: 10.1126/sciadv.adq5484. 
            Epub 2025 Apr 9.
  PUBMED    40203091
REFERENCE   2  (bases 1 to 11481)
  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: "Sci Adv."; 
            date: "2025-04-11"; volume: "11(15):eadq5484. doi"; pages: " 
            10.1126/sciadv.adq5484. Epub 2025 Apr 9"
COMMENT     Sequence Label: PB-TRE3G-dN40-TP53-CCRB
FEATURES             Location/Qualifiers
     source          1..11481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1418..1436
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1454..1472
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1490..1508
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1526..1544
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1562..1580
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1598..1616
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1634..1652
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1697..1721
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    2916..3504
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2969..2989)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3387..3398)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3537..3658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3574..3593)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3628..3647
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     enhancer        3737..4040
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          4318..5334
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5258..5280
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5342..5361
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      5401..5410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5413..5433
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5437..5457
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5461..5481
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5497..5499,5500..5502,5503..6213)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(5542..5563)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     6150..6171
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6223..6279
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6280..7026
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    join(7043..7617,7618..7620,7621..7629)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7210..7227)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7437..7456
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7630..8028
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(8066..8083)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8072..8296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8935..8969
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9460..9476)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9460..9476)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9473..9495)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9484..9500
                     /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(9508..9514,9515..9532,9533..9538))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9553..9574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9691..9708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9862..10450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9942..9961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10621..11412,11413..11481))
                     /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     11244..11263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggcccgat gaggcccttt cgtcttcact
     1381 cgaggatcgc tgatatctcc accccacagt ggtttactcc ctatcagtga tagagaacgt
     1441 atgaagagtt tactccctat cagtgataga gaacgtatgc agactttact ccctatcagt
     1501 gatagagaac gtataaggag tttactccct atcagtgata gagaacgtat gaccagttta
     1561 ctccctatca gtgatagaga acgtatctac agtttactcc ctatcagtga tagagaacgt
     1621 atatccagtt tactccctat cagtgataga gaacgtataa gctttaggcg tgtacggtgg
     1681 gcgcctataa aagcagagct cgtttagtga accgtcagat cgcctggagc aattccacaa
     1741 cacttttgtc ttataccaac tttccgtacc acttcctacc ctcgtaaact taaggttagg
     1801 tggtacggga attcaccatg gatgatttga tgctgtcccc ggacgatatt gaacaatggt
     1861 tcactgaaga cccaggtcca gatgaagctc ccagaatgcc agaggctgct ccccgcgtgg
     1921 cccctgcacc agcagctcct acaccggcgg cccctgcacc agccccctcc tggcccctgt
     1981 catcttctgt cccttcccag aaaacctacc agggcagcta cggtttccgt ctgggcttct
     2041 tgcattctgg gacagccaag tctgtgactt gcacgtactc ccctgccctc aacaagatgt
     2101 tttgccaact ggccaagacc tgccctgtgc agctgtgggt tgattccaca cccccgcccg
     2161 gcacccgcgt ccgcgccatg gccatctaca agcagtcaca gcacatgacg gaggttgtga
     2221 ggcgctgccc ccaccatgag cgctgctcag atagcgatgg tctggcccct cctcagcatc
     2281 ttatccgagt ggaaggaaat ttgcgtgtgg agtatttgga tgacagaaac acttttcgac
     2341 atagtgtggt ggtgccctat gagccgcctg aggttggctc tgactgtacc accatccact
     2401 acaactacat gtgtaacagt tcctgcatgg gcggcatgaa ccggaggccc atcctcacca
     2461 tcatcacact ggaagactcc agtggtaatc tactgggacg gaacagcttt gaggtgcgtg
     2521 tttgtgcctg tcctgggaga gaccggcgca cagaggaaga gaatctccgc aagaaagggg
     2581 agcctcacca cgagctgccc ccagggagca ctaagcgagc actgcccaac aacaccagct
     2641 cctctcccca gccaaagaag aaaccactgg atggagaata tttcaccctt cagatccgtg
     2701 ggcgtgagcg cttcgagatg ttccgagagc tgaatgaggc cttggaactc aaggatgccc
     2761 aggctgggaa ggagccaggg gggagcaggg ctcactccag ccacctgaag tccaaaaagg
     2821 gtcagtctac ctcccgccat aaaaaactca tgttcaagac agaagggcct gactcagact
     2881 gataagatat cgtatacgat ccattaggcg cggccaatca acctctggat tacaaaattt
     2941 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3001 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3061 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3121 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3181 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3241 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3301 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     3361 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3421 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3481 tctccctttg ggccgcctcc ccgcaccggt aaaggccgca atcgatcgcc caccccaact
     3541 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3601 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3661 atgtctgtat acgcactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     3721 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     3781 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     3841 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     3901 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     3961 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4021 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     4081 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     4141 tgggggcggg gggggggggg gcgcgcgcca ggcggggsgg ggsggggsgr ggggsggggs
     4201 ggggsgrggs ggrgaggkgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
     4261 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
     4321 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
     4381 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     4441 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
     4501 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
     4561 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
     4621 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     4681 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     4741 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     4801 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     4861 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     4921 ccgcctcggg ccggggrggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     4981 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     5041 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     5101 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     5161 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     5221 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     5281 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     5341 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattccc ttaaggttac
     5401 gccaccatgg tgccaaagaa gaagaggaaa gtggacccca agaaaaaaag aaaggttgat
     5461 ccaaaaaaga aaagaaaagt acacgtgtca acgcgtatgg tgagcaaggg cgaggagctg
     5521 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     5581 agcgtccgcg gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc
     5641 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc
     5701 gtggcctgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     5761 atgcccgaag gctacgtcca ggagcgcacc atctctttca aggacgacgg tacctacaag
     5821 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     5881 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc
     5941 cacaacgtct atatcacggc cgacaagcag aagaacggca tcaaggctaa cttcaagatc
     6001 cgccacaacg ttgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     6061 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagccatca gtccgccctg
     6121 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     6181 gggattacac atggcatgga cgagctgtac aagggatctg gcgccacgaa cttctctctg
     6241 ttaaaacaag caggagacgt ggaagaaaat cccggtccga tgagccggct ggacaagagc
     6301 aaagtgatca acagcgccct ggaactgctg aacggcgtgg gcatcgaggg cctgaccacc
     6361 cggaagctgg cccagaaact gggcgtggaa cagcccaccc tgtactggca cgtgaagaac
     6421 aagcgggccc tgctggacgc cctgcccatc gagatgctgg accggcacca cacccacagc
     6481 tgccctctgg aaggcgagag ctggcaggac ttcctgcgga acaacgccaa gagctacaga
     6541 tgcgccctgc tgagccaccg ggacggcgcc aaagtgcacc tgggcaccag acccaccgag
     6601 aagcagtacg agacactgga aaaccagctg gccttcctgt gccagcaggg cttcagcctg
     6661 gaaaacgccc tgtacgccct gagcgccgtg ggccacttca ccctgggctg cgtgctggaa
     6721 gaacaggaac accaggtcgc caaagaggaa agagagacac ccaccaccga cagcatgccc
     6781 cccctgctga agcaggccat cgagctgttc gacagacagg gcgccgagcc cgccttcctg
     6841 ttcggcctgg aactgatcat ctgcggcctc gagaagcagc tgaagtgcga gagcggcgga
     6901 cccaccgacg ccctggacga cttcgacctg gacatgctgc ccgccgatgc cctggatgat
     6961 tttgatctgg atatgctccc tgctgacgct ctcgatgact tcgatctcga tatgctgcca
     7021 ggctaagtta gatctaattc cgcccctctc cctccccccc ccctaacgtt actggccgaa
     7081 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     7141 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     7201 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     7261 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     7321 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     7381 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     7441 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     7501 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     7561 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     7621 gccacaacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     7681 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct
     7741 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca
     7801 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     7861 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt
     7921 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     7981 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagt cgagcctcta
     8041 gagttatcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     8101 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     8161 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     8221 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     8281 gcggtgggct ctatggcttc tgaggcggaa gtcaaccagg ttgcagggtt tcctgtcctc
     8341 acaggaacga agtccctaaa gaaacagtgg cagccaggtt tagccccgga attgactgga
     8401 ttcctttttt agggcccatt ggtatggctt tttccccgta tccccccagg tgtctgcagg
     8461 ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccc
     8521 gggctgtccc cgcacgctgc cggctcgggg atgcgggggg agcgccggac cggagcggag
     8581 ccccgggcgg ctcgctgctg ccccctagcg ggggagggac gtaattacat ccctgggggc
     8641 tttggggggg ggctgtccct gatatctata acaagaaaat atatatataa taagttatca
     8701 cgtaagtaga acatgaaata acaatataat tatcgtatga gttaaatctt aaaagtcacg
     8761 taaaagataa tcatgcgtca ttttgactca cgcggtcgtt atagttcaaa atcagtgaca
     8821 cttaccgcat tgacaagcac gcctcacggg agctccaagc ggcgactgag atgtcctaaa
     8881 tgcacagcga cggattcgcg ctatttagaa agagagagca atatttcaag aatgcatgcg
     8941 tcaattttac gcagactatc tttctagggt taatctagct gcatcaggat catatcgtcg
     9001 ggtctttttt ccggctcagt catcgcccaa gctggcgcta tctgggcatc ggggaggaag
     9061 aagcccgtgc cttttcccgc gaggttgaag cggcatggaa agagtttgcc gaggatgact
     9121 gctgctgcat tgacgttgag cgaaaacgca cgtttaccat gatgattcgg gaaggtgtgg
     9181 ccatgcacgc ctttaacggt gaactgttcg ttcaggccac ctgggatacc agttcgtcgc
     9241 ggcttttccg gacacagttc cggatggtca gcccgaagcg catcagcaac ccgaacaata
     9301 ccggcgacag ccggaactgc cgtgccggtg tgcagattaa tgacagcggt gcggcgctgg
     9361 gatattacgt cagcgaggac gggtatcctg gctggatgcc gcagaaatgg acatggatac
     9421 cccgtgagtt acccggcggg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     9481 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     9541 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     9601 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     9661 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     9721 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     9781 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     9841 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     9901 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     9961 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
    10021 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
    10081 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
    10141 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
    10201 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
    10261 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
    10321 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
    10381 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
    10441 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
    10501 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
    10561 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
    10621 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
    10681 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
    10741 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
    10801 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
    10861 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
    10921 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
    10981 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
    11041 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
    11101 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
    11161 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
    11221 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
    11281 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
    11341 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
    11401 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
    11461 acggaaatgt tgaatactca t
//
