Plasmid Name = SFG-TRE-IRTE Database Index Number = 1492 Created by: HIHG Description: FEATURES Locations/Qualifiers plasmid 1..8252 /note="SFG-TRE-IRTE, 8252 bp. Made via ligation #737." misc_feature 2..35 /note="Oligos: #1104, Blunt(Blunt)--BamH I(5') 34 bp." misc_feature 36..8253 /note="pSFGtcLuc+IRTE-: #1105, BamH I(5')--Blunt(Blunt) 8218 bp." CDS 3505..4362 /label="bla" /note="B-lactamase, confers ampicillin resistance, no Pst I." misc_feature 5486..5502 /label="lacO" /note="lac operator; bound by LacI repressor to repress lac operon expression" misc_feature 7818..7853 /label="tetO" /note="tetracycline operator from Transposon Tn10, used in Bujard plasmids for tet regulation." misc_feature 7860..7895 /label="tetO" /note="tetracycline operator from Transposon Tn10, used in Bujard plasmids for tet regulation." misc_feature 7902..7937 /label="tetO" /note="tetracycline operator from Transposon Tn10, used in Bujard plasmids for tet regulation." misc_feature 7944..7979 /label="tetO" /note="tetracycline operator from Transposon Tn10, used in Bujard plasmids for tet regulation." misc_feature 7986..8021 /label="tetO" /note="tetracycline operator from Transposon Tn10, used in Bujard plasmids for tet regulation." misc_feature 8028..8063 /label="tetO" /note="tetracycline operator from Transposon Tn10, used in Bujard plasmids for tet regulation." misc_feature 8070..8105 /label="tetO" /note="tetracycline operator from Transposon Tn10, used in Bujard plasmids for tet regulation." misc_feature 4661..4704 /label="Low_copy" /note="Tag for low copy plasmid replication xxpBR322xx" misc_feature 8130..8181 /label="CMV_TATA" /note="CMV TATA box sequences, from -52 to -1. Bujard version, from UHC 13-3, with Sac I site." repeat_region 2068..2135 /label="R_MLV" /note="R region fro Moloney Murine Leukemia Virus." repeat_region 6384..6451 /label="R_MLV" /note="R region fro Moloney Murine Leukemia Virus." repeat_region 2136..2210 /label="U5_MLV" /note="U5 region of Moloney MLV." repeat_region 6452..6526 /label="U5_MLV" /note="U5 region of Moloney MLV." CDS 625..1629 /label="rtTA" /note="reverse tet transactivator, from Bujard." misc_feature 1788..1800 /label="MLV_U3_tip" /note="Tip of both MLV and MPSV U3. Begins the U3 region." misc_feature 5935..5947 /label="MLV_U3_tip" /note="Tip of both MLV and MPSV U3. Begins the U3 region." CDS 2910..3068 /label="lacZ_alphaxx" /note="Fragment of lacZ alpha sequence that is used in pUC. Should also pick up the fragment in other plasmids." misc_feature 7004..7021 /label="SFG_gag" /note="gag ATG, as it appears in SFG constructs." misc_feature 6794..6811 /label="MFGS_csStop" /note="Engineered stop codon from MFG-S, SFG, and MMP vector constructs. Terminates the cell surface gag orf 18 amino acids after the CTG start codon." misc_feature 6740..7003 /label="MFGS_CSgag" /note="Mutated form of the Moloney Cell Surface gag extension. Starting at the CTG, this sequence is translated into the gag open reading frame, in order to make the cell surface form of the gag polyprotein xxglycogagxx. Here there are two mutations, one which introduces a stop codon after the 18th codon, and another which is compensatory for maintaining the secondary structure of the packaging signal in this region." intron 6589..7529 /label="MoMLV_intron" /note="Intron from SFG. Note Nhe I site after gag atg." misc_feature complement(6516..6528) /label="MLV_U3_tip" /note="Tip of both MLV and MPSV U3. Begins the U3 region." misc_feature complement(6529..6546) /label="PBS-P" /note="MoMLV tRNA primer binding site, for proline tRNA." misc_feature 7808..7838 /label="Correction-114" /note="SFG-TRE-IRTE constructs, needs more sequencing to confirm if this ever really was an error, as opposed to misaligned repeats. Makes the first tetO repeat the same as the fifth. This sequence has been found to be erroneous for some plasmids in our laboratory. It would be prudent to sequence the DNA to confirm the accuracy of your records. The sequence specified by this feature was actually found to be: AGCCCTCGAG TTTACCAGTC CCTATCAGTG A. " misc_feature 5512..5535 /label="Correction-189" /note="pUC18 instability. This deletion has been shown to occur in many plasmids based upon pUC18 and similar plasmids. It involves a deletion of the cytosine nucleotide from the second codon of the lacZ alpha gene, just upstream of the polylinker. Some constructs in our lab have the deletion, and some do not. There is no apparent restriction enzyme polymorphism, but the deletion can affect the performance of some sequencing primers. See Lobet, Y. et al., 1989, NAR v17, 4897. This sequence has been found to be erroneous for some plasmids in our laboratory. It would be prudent to sequence the DNA to confirm the accuracy of your records. The sequence specified by this feature was actually found to be: GGAAACAGCT ATGACATGAT TAC. " misc_feature complement(4668..4699) /label="Correction-190" /note="Difference between low and high copy plasmids. Many of our lab plasmids are high copy, yet the sequence file suggests that they are low copy. This correction could be applied to make the sequence properly represent the actual DNA sequence for a high copy plasmid. It was confirmed in the full plasmid sequencing of CMMP-MR1-1TcR, which has the pUC18 backbone. This sequence has been found to be erroneous for some plasmids in our laboratory. It would be prudent to sequence the DNA to confirm the accuracy of your records. The sequence specified by this feature was actually found to be: ACGGCTACAC TAGAAGAACA GTATTTGGTA TC. " CDS 8194..8217 /label="HGTI-761_Max=" /note="PCR or sequencing primer located in CMV sequence just after 5xx end of RNA transcript. Max recorded sequencing read = bases. Perfect match Tm = 63.9 C, although only 17 nt are required to match. The actual sequence of the primer is GAGACGCCAT CCACGCTGTT TTGA." CDS 7107..7131 /label="HGTI-1043_Max=0" /note="Sequencing primers for MLV gagpol. Max recorded sequencing read = 0 bases. Perfect match Tm = 61.1 C, although only 17 nt are required to match. The actual sequence of the primer is GACGTTGGGT TACCTTCTGC TCTGC." CDS 7307..7335 /label="HGTI-1044_Max=561" /note="Sequencing primers for MLV gagpol. Max recorded sequencing read = 561 bases. Perfect match Tm = 67.0 C, although only 17 nt are required to match. The actual sequence of the primer is TTTGTACACC CTAAGCCTCC GCCTCCTCT." CDS 7460..7489 /label="HGTI-1065_Max=679" /note="Sequencing primers for MLV gagpol. Max recorded sequencing read = 679 bases. Perfect match Tm = 73.6 C, although only 17 nt are required to match. The actual sequence of the primer is GCACCCCCGC CCCTTGTAAA CTTCCCTGAC ." CDS 7653..7678 /label="HGTI-1066_Max=663" /note="Sequencing primers for MLV gagpol. Max recorded sequencing read = 663 bases. Perfect match Tm = 62.8 C, although only 17 nt are required to match. The actual sequence of the primer is TCCGCCGACA CCAGACTAAG AACCTA." CDS complement(7046..7074) /label="HGTI-1067_Max=524" /note="Sequencing primers for MLV gagpol. Max recorded sequencing read = 524 bases. Perfect match Tm = 65.4 C, although only 17 nt are required to match. The actual sequence of the primer is ATCCGCTCGA CATCTTTCCA GTGACCTAA." CDS complement(7251..7278) /label="HGTI-1068_Max=458" /note="Sequencing primers for MLV gagpol. Max recorded sequencing read = 458 bases. Perfect match Tm = 69.3 C, although only 17 nt are required to match. The actual sequence of the primer is GCCAAGGCTT CCCAGGTCAC GATGTAGG." CDS complement(7399..7424) /label="HGTI-1069_Max=464" /note="Sequencing primers for MLV gagpol. Max recorded sequencing read = 464 bases. Perfect match Tm = 57.5 C, although only 17 nt are required to match. The actual sequence of the primer is CTAGAGAAGG AGTGAGGGCT GGATAA." CDS complement(7592..7613) /label="HGTI-1090_Max=447" /note="Sequencing primers for MLV gagpol. Max recorded sequencing read = 447 bases. Perfect match Tm = 60.2 C, although only 17 nt are required to match. The actual sequence of the primer is CCGGTCGGTC CAGTTGTTCT TG." CDS complement(8164..8205) /label="HGTI-1192_Max=" /note="linker from SacI of CMV bridging into BglII of HIV R. Max recorded sequencing read = bases. Perfect match Tm = 74.8 C, although only 17 nt are required to match. The actual sequence of the primer is gatcTGGTCT AACCAGAGAG ACCCcggttc actaaacgAG CT." CDS 3017..3042 /label="HGTI-1288_Max=" /note="sequencing primers for AAV backbone Max recorded sequencing read = bases. Perfect match Tm = 72.8 C, although only 17 nt are required to match. The actual sequence of the primer is CCGCACCGAT CGCCCTTCCC AACAGT." CDS complement(3171..3194) /label="HGTI-1291_Max=577" /note="sequencing primers for AAV backbone Max recorded sequencing read = 577 bases. Perfect match Tm = 71.4 C, although only 17 nt are required to match. The actual sequence of the primer is GTCAGCGGGT GTTGGCGGGT GTCG." CDS 7732..7755 /label="HGTI-1292_Max=563" /note="retrovirus sequencing primer forward Max recorded sequencing read = 563 bases. Perfect match Tm = 61.8 C, although only 17 nt are required to match. The actual sequence of the primer is caaagtagac ggcatcgcag cttg." CDS complement(1690..1715) /label="HGTI-1293_Max=467" /note="sequencing primer for retrovirus reverse Max recorded sequencing read = 467 bases. Perfect match Tm = 51.1 C, although only 17 nt are required to match. The actual sequence of the primer is tgatattgtt gagtcaaaac tagagc." CDS 6776..6805 /label="HGTI-1605_Max=446" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = 446 bases. Perfect match Tm = 70.6 C, although only 17 nt are required to match. The actual sequence of the primer is GTGGCCCGAC CTGAGTCCTA AAATCCCGAT ." CDS 7026..7055 /label="HGTI-1606_Max=559" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = 559 bases. Perfect match Tm = 61.3 C, although only 17 nt are required to match. The actual sequence of the primer is CCACTCCCTT AAGTTTGACC TTAGGTCACT ." CDS 7276..7305 /label="HGTI-1607_Max=163" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = 163 bases. Perfect match Tm = 75.7 C, although only 17 nt are required to match. The actual sequence of the primer is GGCTTTTGAC CCCCCTCCCT GGGTCAAGCC ." CDS 7526..7555 /label="HGTI-1608_Max=716" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = 716 bases. Perfect match Tm = 57.3 C, although only 17 nt are required to match. The actual sequence of the primer is CAAGCTCACT TACAGGCTCT CTACTTAGTC ." CDS 7776..7805 /label="HGTI-1609_Max=0" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = 0 bases. Perfect match Tm = 80.3 C, although only 17 nt are required to match. The actual sequence of the primer is AGGCTGCCGA CCCCGGGGGT GGACCATCCT ." CDS 1734..1763 /label="HGTI-1627_Max=509" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = 509 bases. Perfect match Tm = 54.4 C, although only 17 nt are required to match. The actual sequence of the primer is GTACGAGCCA TAGATAAAAT AAAAGATTTT ." CDS complement(1760..1789) /label="HGTI-1628_Max=" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = bases. Perfect match Tm = 62.2 C, although only 17 nt are required to match. The actual sequence of the primer is ttcccccctt tttctggaga ctaaataaaa ." CDS complement(7552..7581) /label="HGTI-1647_Max=607" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = 607 bases. Perfect match Tm = 69.6 C, although only 17 nt are required to match. The actual sequence of the primer is gccagaggtc tccagacttc gtgctggact ." CDS complement(7302..7331) /label="HGTI-1648_Max=" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = bases. Perfect match Tm = 70.1 C, although only 17 nt are required to match. The actual sequence of the primer is gaggcggagg cttagggtgt acaaagggct ." CDS complement(7052..7081) /label="HGTI-1649_Max=" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = bases. Perfect match Tm = 71 C, although only 17 nt are required to match. The actual sequence of the primer is gtgagcgatc cgctcgacat ctttccagtg ." CDS complement(6802..6831) /label="HGTI-1650_Max=130" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = 130 bases. Perfect match Tm = 70.3 C, although only 17 nt are required to match. The actual sequence of the primer is gggggtgcac caaagagtcc taaacgatcg ." CDS complement(6553..6582) /label="HGTI-1651_Max=" /note="Sequencing primers for MMP-FAA, from LTR to LTR Max recorded sequencing read = bases. Perfect match Tm = 82.9 C, although only 17 nt are required to match. The actual sequence of the primer is cgggggggtc ggtggtccct gggcaggggt ." CDS complement(3135..3165) /label="HGTI-1662_Max=680" /note="Series of primers for construction of 4 variations of Adenovirus helper constructs, with potential for deletions in Psi. Max recorded sequencing read = 680 bases. Perfect match Tm = 67.2 C, although only 17 nt are required to match. The actual sequence of the primer is GGCTTAACTA TGCGGCATCA GAGCAGATTG T." CDS 2905..2927 /label="HGTI-1762_Max=" /note="Sequencing primer in lacZ, at 5xx end and facing upstream. Max recorded sequencing read = bases. Perfect match Tm = 57.4 C, although only 17 nt are required to match. The actual sequence of the primer is attcactggc cgtcgtttta caa." CDS 5505..5526 /label="HGTI-1805_Max=673" /note="std sequencing primer -47 Max recorded sequencing read = 673 bases. Perfect match Tm = 49.4 C, although only 17 nt are required to match. The actual sequence of the primer is tcacacagga aacagctatg ac." CDS 49..68 /label="HGTI-1839_Max=" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = bases. Perfect match Tm = 58.4 C, although only 17 nt are required to match. The actual sequence of the primer is tccccccccc ctaacgttac ." CDS 349..375 /label="HGTI-1840_Max=418" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 418 bases. Perfect match Tm = 59.1 C, although only 17 nt are required to match. The actual sequence of the primer is aagccacgtg tataagatac acctgca." CDS complement(5129..5148) /label="HGTI-1850_Max=589" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 589 bases. Perfect match Tm = 59.7 C, although only 17 nt are required to match. The actual sequence of the primer is taaaaaggcc gcgttgctgg ." CDS complement(4830..4848) /label="HGTI-1851_Max=601" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 601 bases. Perfect match Tm = 60.1 C, although only 17 nt are required to match. The actual sequence of the primer is cgaccgctgc gccttatcc." CDS complement(4519..4548) /label="HGTI-1852_Max=372" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 372 bases. Perfect match Tm = 58.8 C, although only 17 nt are required to match. The actual sequence of the primer is aaaaggatct caagaagatc ctttgatctt ." CDS complement(4227..4248) /label="HGTI-1853_Max=0" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 0 bases. Perfect match Tm = 61.0 C, although only 17 nt are required to match. The actual sequence of the primer is ccccagtgct gcaatgatac cg." CDS complement(3928..3948) /label="HGTI-1854_Max=650" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 650 bases. Perfect match Tm = 58.4 C, although only 17 nt are required to match. The actual sequence of the primer is agcggttagc tccttcggtc c." CDS complement(3625..3648) /label="HGTI-1855_Max=661" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 661 bases. Perfect match Tm = 58.0 C, although only 17 nt are required to match. The actual sequence of the primer is atccagttcg atgtaaccca ctcg." CDS complement(3318..3348) /label="HGTI-1856_Max=711" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 711 bases. Perfect match Tm = 58.3 C, although only 17 nt are required to match. The actual sequence of the primer is gacattaacc tataaaaata ggcgtatcac g." CDS complement(141..158) /label="HGTI-1870_Max=52" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Also good for IRES upstream junctions. Max recorded sequencing read = 52 bases. Perfect match Tm = 59.8 C, although only 17 nt are required to match. The actual sequence of the primer is cgggccctca cattgcca." CDS complement(434..458) /label="HGTI-1871_Max=499" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 499 bases. Perfect match Tm = 59.7 C, although only 17 nt are required to match. The actual sequence of the primer is ttgttgaata cgcttgagga gagcc." CDS 5339..5361 /label="HGTI-1880_Max=690" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 690 bases. Perfect match Tm = 59.4 C, although only 17 nt are required to match. The actual sequence of the primer is gttggccgat tcattaatgc agc." CDS 5039..5064 /label="HGTI-1881_Max=511" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 511 bases. Perfect match Tm = 59 C, although only 17 nt are required to match. The actual sequence of the primer is atctttatag tcctgtcggg tttcgc." CDS 4739..4766 /label="HGTI-1882_Max=565" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 565 bases. Perfect match Tm = 60.5 C, although only 17 nt are required to match. The actual sequence of the primer is catacctcgc tctgctaatc ctgttacc." CDS 4439..4467 /label="HGTI-1883_Max=564" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 564 bases. Perfect match Tm = 58.2 C, although only 17 nt are required to match. The actual sequence of the primer is tgaagatcct ttttgataat ctcatgacc." CDS 4139..4163 /label="HGTI-1884_Max=593" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 593 bases. Perfect match Tm = 59.6 C, although only 17 nt are required to match. The actual sequence of the primer is ataaagttgc aggaccactt ctgcg." CDS 3839..3866 /label="HGTI-1885_Max=654" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 654 bases. Perfect match Tm = 60.0 C, although only 17 nt are required to match. The actual sequence of the primer is cggatggcat gacagtaaga gaattatg." CDS 3539..3559 /label="HGTI-1886_Max=647" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 647 bases. Perfect match Tm = 60.2 C, although only 17 nt are required to match. The actual sequence of the primer is ccttttttgc ggcattttgc c." CDS 3238..3257 /label="HGTI-1887_Max=206" /note="Full plasmid sequencing for Add-BGFP-FasL-Ires-FADD Max recorded sequencing read = 206 bases. Perfect match Tm = 58.6 C, although only 17 nt are required to match. The actual sequence of the primer is aagctgtgac cgtctccggg ." CDS complement(6950..6977) /label="HGTI-1979_Max=0" /note="MLV LTR sequencing Max recorded sequencing read = 0 bases. Perfect match Tm = 61.3 C, although only 17 nt are required to match. The actual sequence of the primer is TCAGACAGAG ACAACACAGA ACGATGCT." CDS complement(6807..6831) /label="HGTI-1980_Max=0" /note="MLV LTR sequencing Max recorded sequencing read = 0 bases. Perfect match Tm = 61.8 C, although only 17 nt are required to match. The actual sequence of the primer is GGGGGTGCAC CAAAGAGTCC TAAAC." CDS 5491..5514 /label="HGTI-2082_Max=0" /note="Reverse sequencing primer for pUC. Purpose of the oligo: Sequencing Max recorded sequencing read = 0 bases. Perfect match Tm = 56.6 C, although only 17 nt are required to match. The actual sequence of the primer is AGCGGATAAC AATTTCACAC AGGA." ^^ GCCTTAAGCAATTGCGGCCGCTACGTAATCGATGGATCCCCCTCTCCCTCCCCCCCCCCTAACGTTACTGGCCGAAGCCGCTTGGAATAAGGCCGGTGTGCGTTTGTCTATATGTTATTTTCCACCATATTGCCGTCTTTTGGCAATGTGAGGGCCCGGAAACCTGGCCCTGTCTTCTTGACGAGCATTCCTAGGGGTCTTTCCCCTCTCGCCAAAGGAATGCAAGGTCTGTTGAATGTCGTGAAGGAAGCAGTTCCTCTGGAAGCTTCTTGAAGACAAACAACGTCTGTAGCGACCCTTTGCAGGCAGCGGAACCCCCCACCTGGCGACAGGTGCCTCTGCGGCCAAAAGCCACGTGTATAAGATACACCTGCAAAGGCGGCACAACCCCAGTGCCACGTTGTGAGTTGGATAGTTGTGGAAAGAGTCAAATGGCTCTCCTCAAGCGTATTCAACAAGGGGCTGAAGGATGCCCAGAAGGTACCCCATTGTATGGGATCTGATCTGGGGCCTCGGTGCACATGCTTTACATGTGTTTAGTCGAGGTTAAAAAACGTCTAGGCCCCCCGAACCACGGGGACGTGGTTTTCCTTTGAAAAACACGATGATAAGCTTGCCACAACCATGTCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTTGGTGTAGAGCAGCCTACACTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAAAAGGGGAAAGCTGGCAAGATTTTTTACGCAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCAATGGAGCAAAAGTACATTCAGATACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAACGCGTTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCCGCGTACAGCCGCGCGCGTACGAAAAACAATTACGGGTCTACCATCGAGGGCCTGCTCGATCTCCCGGACGACGACGCCCCCGAAGAGGCGGGGCTGGCGGCTCCGCGCCTGTCCTTTCTCCCCGCGGGACACACGCGCAGACTGTCGACGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGTAGGGGGCGCGAAGATCCGGATTAGTCCAATTTGTTAAAGACAGGATATCAGTGGTCCAGGCTCTAGTTTTGACTCAACAATATCACCAGCTGAAGCCTATAGAGTACGAGCCATAGATAAAATAAAAGATTTTATTTAGTCTCCAGAAAAAGGGGGGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATACATAACTGAGAATAGAGAAGTTCAGATCAAGGTCAGGAACAGATGGAACAGCTCGACCCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTCACACATGCAGCATGTATCAAAATTAATTTGGTTTTTTTTCTTAAGTATTTACATTAAATGGCCATAGTACTTAAAGTTACATTGGCTTCCTTGAAATAAACATGGAGTATTCAGAATGTGTCATAAATATTTCTAATTTTAAGATAGTATCTCCATTGGCTTTCTACTTTTTCTTTTATTTTTTTTTGTCCTCTGTCTTCCATTTGTTGTTGTTGTTGTTTGTTTGTTTGTTTGTTGGTTGGTTGGTTAATTTTTTTTTAAAGATCCTACACTATAGTTCAAGCTAGACTATTAGCTACTCTGTAACCCAGGGTGACCTTGAAGTCATGGGTAGCCTGCTGTTTTAGCCTTCCCACATCTAAGATTACAGGTATGAGCTATCATTTTTGGTATATTGATTGATTGATTGATTGATGTGTGTGTGTGTGATTGTGTTTGTGTGTGTGACTGTGAAAATGTGTGTATGGGTGTGTGTGAATGTGTGTATGTATGTGTGTGTGTGAGTGTGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGACTGTGTCTATGTGTATGACTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTTGTGAAAAAATATTCTATGGTAGTGAGAGCCAACGCTCCGGCTCAGGTGTCAGGTTGGTTTTTGAGACAGAGTCTTTCACTTAGCTTGGAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGATGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTTGCTCTTAGGAGTTTCCTAATACATCCCAAACTCAAATATATAAAGCATTTGACTTGTTCTATGCCCTAGGGGGCGGGGGGAAGCTAAGCCAGCTTTTTTTAACATTTAAAATGTTAATTCCATTTTAAATGCACAGATGTTTTTATTTCATAAGGGTTTCAATGTGCATGAATGCTGCAATATTCCTGTTACCAAAGCTAGTATAAATAAAAATAGATAAACGTGGAAATTACTTAGAGTTTCTGTCATTAACGTTTCCTTCCTCAGTTGACAACATAAATGCGCTGCTGAGCAAGCCAGTTTGCATCTGTCAGGATCAATTTCCCATTATGCCAGTCATATTAATTACTAGTCAATTAGTTGATTTTTATTTTTGACATATACATGTGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATACATAACTGAGAATAGAGAAGTTCAGATCAAGGTCAGGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTATGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGACCCCTGCCCAGGGACCACCGACCCACCACCGGGAGGTAAGCTGGCCAGCAACTTATCTGTGTCTGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAACTAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCGGAACACCCGGCCGCAACCCTGGGAGACGTCCCAGGGACTTCGGGGGCCGTTTTTGTGGCCCGACCTGAGTCCTAAAATCCCGATCGTTTAGGACTCTTTGGTGCACCCCCCTTAGAGGAGGGATATGTGGTTCTGGTAGGAGACGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGGACCGAAGCCGCGCCGCGCGTCTTGTCTGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTGTATTTGTCTGAAAATATGGGCCCGGGCTAGCCTGTTACCACTCCCTTAAGTTTGACCTTAGGTCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTTGGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACCTTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACACCCAGACCAGGTCCCCTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGGTCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCCCTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACTCCTTCTCTAGGCGCCCCCATATGGCCATATGAGATCTTATATGGGGCACCCCCGCCCCTTGTAAACTTCCCTGACCCTGACATGACAAGAGTTACTAACAGCCCCTCTCTCCAAGCTCACTTACAGGCTCTCTACTTAGTCCAGCACGAAGTCTGGAGACCTCTGGCGGCAGCCTACCAAGAACAACTGGACCGACCGGTGGTACCTCACCCTTACCGAGTCGGCGACACAGTGTGGGTCCGCCGACACCAGACTAAGAACCTAGAACCTCGCTGGAAAGGACCTTACACAGTCCTGCTGACCACCCCCACCGCCCTCAAAGTAGACGGCATCGCAGCTTGGATACACGCCGCCCACGTGAAGGCTGCCGACCCCGGGGGTGGACCATCCTCTAGCCCTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGCTCGGTACCCGGGTCGAGGTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACCTCCATAGAAGACACCGGGACCGATCCGACCTGC