LOCUS       sequence-473536-       11598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A piggybac-based cloning vector containing TRE3G promoter-driven 
            full-length 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 11598)
  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 11598)
  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-TP53-CCRB
FEATURES             Location/Qualifiers
     source          1..11598
                     /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"
     CDS             1818..2999
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML
                     SPDDIEQWFTEDPGPDEAPRMPEAAPRVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     misc_feature    3033..3621
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3086..3106)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3504..3515)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3654..3775
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3691..3710)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3745..3764
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     enhancer        3854..4157
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          4435..5451
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5375..5397
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5459..5478
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      5518..5527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5530..5550
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5554..5574
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5578..5598
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5614..5616,5617..5619,5620..6330)
                     /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(5659..5680)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     6267..6288
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6340..6396
                     /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             6397..7143
                     /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(7160..7734,7735..7737,7738..7746)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7327..7344)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7554..7573
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7747..8145
                     /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(8183..8200)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8189..8413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   9052..9086
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9577..9593)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9577..9593)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9590..9612)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9601..9617
                     /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(9625..9631,9632..9649,9650..9655))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9670..9691
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9808..9825)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9979..10567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10059..10078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10738..11529,11530..11598))
                     /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     11361..11380
                     /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 gaggagccgc agtcagatcc tagcgtcgag ccccctctga
     1861 gtcaggaaac attttcagac ctatggaaac tacttcctga aaacaacgtt ctgtccccct
     1921 tgccgtccca agcaatggat gatttgatgc tgtccccgga cgatattgaa caatggttca
     1981 ctgaagaccc aggtccagat gaagctccca gaatgccaga ggctgctccc cgcgtggccc
     2041 ctgcaccagc agctcctaca ccggcggccc ctgcaccagc cccctcctgg cccctgtcat
     2101 cttctgtccc ttcccagaaa acctaccagg gcagctacgg tttccgtctg ggcttcttgc
     2161 attctgggac agccaagtct gtgacttgca cgtactcccc tgccctcaac aagatgtttt
     2221 gccaactggc caagacctgc cctgtgcagc tgtgggttga ttccacaccc ccgcccggca
     2281 cccgcgtccg cgccatggcc atctacaagc agtcacagca catgacggag gttgtgaggc
     2341 gctgccccca ccatgagcgc tgctcagata gcgatggtct ggcccctcct cagcatctta
     2401 tccgagtgga aggaaatttg cgtgtggagt atttggatga cagaaacact tttcgacata
     2461 gtgtggtggt gccctatgag ccgcctgagg ttggctctga ctgtaccacc atccactaca
     2521 actacatgtg taacagttcc tgcatgggcg gcatgaaccg gaggcccatc ctcaccatca
     2581 tcacactgga agactccagt ggtaatctac tgggacggaa cagctttgag gtgcgtgttt
     2641 gtgcctgtcc tgggagagac cggcgcacag aggaagagaa tctccgcaag aaaggggagc
     2701 ctcaccacga gctgccccca gggagcacta agcgagcact gcccaacaac accagctcct
     2761 ctccccagcc aaagaagaaa ccactggatg gagaatattt cacccttcag atccgtgggc
     2821 gtgagcgctt cgagatgttc cgagagctga atgaggcctt ggaactcaag gatgcccagg
     2881 ctgggaagga gccagggggg agcagggctc actccagcca cctgaagtcc aaaaagggtc
     2941 agtctacctc ccgccataaa aaactcatgt tcaagacaga agggcctgac tcagactgat
     3001 aagatatcgt atacgatcca ttaggcgcgg ccaatcaacc tctggattac aaaatttgtg
     3061 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3121 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3181 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3241 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3301 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3361 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3421 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3481 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3541 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3601 ccctttgggc cgcctccccg caccggtaaa ggccgcaatc gatcgcccac cccaacttgt
     3661 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3721 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3781 tctgtatacg cactagttat taatagtaat caattacggg gtcattagtt catagcccat
     3841 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     3901 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     3961 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4021 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4081 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4141 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     4201 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     4261 gggcgggggg ggggggggcg cgcgccaggc ggggsggggs ggggsgrggg gsggggsggg
     4321 gsgrggsggr gaggkgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     4381 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     4441 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
     4501 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
     4561 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
     4621 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt
     4681 gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg
     4741 cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt
     4801 gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     4861 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     4921 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     4981 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     5041 cctcgggccg gggrgggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     5101 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     5161 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     5221 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     5281 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     5341 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     5401 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     5461 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattccctta aggttacgcc
     5521 accatggtgc caaagaagaa gaggaaagtg gaccccaaga aaaaaagaaa ggttgatcca
     5581 aaaaagaaaa gaaaagtaca cgtgtcaacg cgtatggtga gcaagggcga ggagctgttc
     5641 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     5701 gtccgcggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc
     5761 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcgtg
     5821 gcctgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     5881 cccgaaggct acgtccagga gcgcaccatc tctttcaagg acgacggtac ctacaagacc
     5941 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     6001 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac
     6061 aacgtctata tcacggccga caagcagaag aacggcatca aggctaactt caagatccgc
     6121 cacaacgttg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     6181 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gccatcagtc cgccctgagc
     6241 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     6301 attacacatg gcatggacga gctgtacaag ggatctggcg ccacgaactt ctctctgtta
     6361 aaacaagcag gagacgtgga agaaaatccc ggtccgatga gccggctgga caagagcaaa
     6421 gtgatcaaca gcgccctgga actgctgaac ggcgtgggca tcgagggcct gaccacccgg
     6481 aagctggccc agaaactggg cgtggaacag cccaccctgt actggcacgt gaagaacaag
     6541 cgggccctgc tggacgccct gcccatcgag atgctggacc ggcaccacac ccacagctgc
     6601 cctctggaag gcgagagctg gcaggacttc ctgcggaaca acgccaagag ctacagatgc
     6661 gccctgctga gccaccggga cggcgccaaa gtgcacctgg gcaccagacc caccgagaag
     6721 cagtacgaga cactggaaaa ccagctggcc ttcctgtgcc agcagggctt cagcctggaa
     6781 aacgccctgt acgccctgag cgccgtgggc cacttcaccc tgggctgcgt gctggaagaa
     6841 caggaacacc aggtcgccaa agaggaaaga gagacaccca ccaccgacag catgcccccc
     6901 ctgctgaagc aggccatcga gctgttcgac agacagggcg ccgagcccgc cttcctgttc
     6961 ggcctggaac tgatcatctg cggcctcgag aagcagctga agtgcgagag cggcggaccc
     7021 accgacgccc tggacgactt cgacctggac atgctgcccg ccgatgccct ggatgatttt
     7081 gatctggata tgctccctgc tgacgctctc gatgacttcg atctcgatat gctgccaggc
     7141 taagttagat ctaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc
     7201 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     7261 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     7321 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     7381 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     7441 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     7501 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     7561 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     7621 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     7681 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     7741 acaaccatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct
     7801 acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc
     7861 gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa
     7921 ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg
     7981 atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc
     8041 gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt
     8101 gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagtcga gcctctagag
     8161 ttatctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     8221 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     8281 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     8341 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     8401 gtgggctcta tggcttctga ggcggaagtc aaccaggttg cagggtttcc tgtcctcaca
     8461 ggaacgaagt ccctaaagaa acagtggcag ccaggtttag ccccggaatt gactggattc
     8521 cttttttagg gcccattggt atggcttttt ccccgtatcc ccccaggtgt ctgcaggctc
     8581 aaagagcagc gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg
     8641 ctgtccccgc acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc
     8701 cgggcggctc gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt
     8761 gggggggggc tgtccctgat atctataaca agaaaatata tatataataa gttatcacgt
     8821 aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa
     8881 aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt
     8941 accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc
     9001 acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat gcatgcgtca
     9061 attttacgca gactatcttt ctagggttaa tctagctgca tcaggatcat atcgtcgggt
     9121 cttttttccg gctcagtcat cgcccaagct ggcgctatct gggcatcggg gaggaagaag
     9181 cccgtgcctt ttcccgcgag gttgaagcgg catggaaaga gtttgccgag gatgactgct
     9241 gctgcattga cgttgagcga aaacgcacgt ttaccatgat gattcgggaa ggtgtggcca
     9301 tgcacgcctt taacggtgaa ctgttcgttc aggccacctg ggataccagt tcgtcgcggc
     9361 ttttccggac acagttccgg atggtcagcc cgaagcgcat cagcaacccg aacaataccg
     9421 gcgacagccg gaactgccgt gccggtgtgc agattaatga cagcggtgcg gcgctgggat
     9481 attacgtcag cgaggacggg tatcctggct ggatgccgca gaaatggaca tggatacccc
     9541 gtgagttacc cggcgggcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     9601 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     9661 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     9721 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     9781 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     9841 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     9901 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     9961 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
    10021 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
    10081 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
    10141 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
    10201 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
    10261 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
    10321 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
    10381 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
    10441 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
    10501 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
    10561 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
    10621 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
    10681 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
    10741 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
    10801 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
    10861 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
    10921 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
    10981 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
    11041 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
    11101 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
    11161 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
    11221 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
    11281 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
    11341 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
    11401 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
    11461 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
    11521 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
    11581 gaaatgttga atactcat
//
