LOCUS Exported 10628 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION doxycycline-inducible expression of human GFI1B (HA tagged) in mammalian cells. ACCESSION . VERSION . KEYWORDS pLV-tetO-HA-GFI1B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10628) AUTHORS Gomes AM, Kurochkin I, Chang B, Daniel M, Law K, Satija N, Lachmann A, Wang Z, Ferreira L, Ma'ayan A, Chen BK, Papatsenko D, Lemischka IR, Moore KA, Pereira CF TITLE Cooperative Transcription Factor Induction Mediates Hemogenic Reprogramming. JOURNAL Cell Rep. 2018 Dec 4;25(10):2821-2835.e7. doi: 10.1016/j.celrep.2018.11.032. PUBMED 30517869 REFERENCE 2 (bases 1 to 10628) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10628 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..315 /label=tight TRE promoter /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 9..27 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 45..63 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 80..98 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 116..134 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 152..170 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 187..205 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 223..241 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 288..312 /label=LNCX /note="Human CMV promoter, forward primer" CDS 337..363 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 1407..1959 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1551..1568) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 1778..1797 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 1971..2690 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1971..2687 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2016..2037) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2277..2296) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2624..2645 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2700..3288 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2753..2773) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3171..3182) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3812..3992 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(4018..4035) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4024..4248 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4294..4722 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4381..4400) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4591..4612 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4731..4751) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4736..5065 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4916..5051 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4978..4997 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 5113..5160 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 5179..5553 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 5683..5804 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5720..5739) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5774..5793 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5853..5869) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5853..5869) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5866..5888) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5877..5893 /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(5901..5931) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5946..5967 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6084..6101) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6255..6843) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6335..6354) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7014..7874) /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 7637..7656 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7875..7979) /gene="bla" /label=AmpR promoter primer_bind complement(8054..8073) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 8245..8624 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 8626..8824 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 8779..8799 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 8842..9022 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 9069..9194 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 9687..9920 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 10105..10149 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 10447..10564 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag 61 agaacgatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct 121 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg 181 agtttatccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag 241 aacgtatgtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg 301 aaccgtcaga tcgcctgact agaattcgcc accatgtacc catacgatgt tccagattac 361 gcttctagag ctagcatgcc acgctccttc ctggtgaaga gcaagaaggc tcacacctac 421 caccagcccc gtgtgcagga agatgaaccg ctctggcctc ctgcccttac cccggtgccc 481 agagaccagg ctccaagcaa cagccctgtc cttagcactc tattcccaaa ccagtgcctg 541 gactggacca acctcaaacg agagccggag ctggagcagg accagaactt ggccaggatg 601 gccccggcac cagagggccc cattgtgctg tcccgacccc aggatgggga ctctccactg 661 tccgactcac ccccattcta caagcctagc ttctcctggg acaccttggc cacaacctat 721 ggccacagct accggcaggc cccctccacc atgcagtcag ccttcctgga gcactccgtc 781 agcctgtacg gcagtcctct tgtgcccagc actgagcccg ccttggactt cagcctccgc 841 tactccccag gcatggatgc gtaccactgt gtgaagtgca acaaggtctt ctccacccct 901 cacgggctcg aagtgcatgt gcgacgctcc catagtggga cccggccctt cgcctgtgac 961 atctgcggca aaaccttcgg ccacgctgtg agcctggagc agcacacgca cgtccactcc 1021 caggagcgca gcttcgagtg ccgcatgtgc ggcaaggcct tcaagcgctc gtccacgctg 1081 tccacccacc tgctcatcca ctcagacacg cggccctacc cctgccagtt ctgcggcaag 1141 cgtttccacc agaagtccga catgaagaag cacacctaca tccacacagg tgagaagccg 1201 cacaagtgcc aggtgtgcgg aaaggccttc agccagagct ccaacctcat cacccacagc 1261 cgcaagcaca caggcttcaa gcccttcagc tgtgagctgt gcaccaaagg cttccagcgc 1321 aaggtggacc tgcggcggca ccgcgagagc cagcacaatc tcaagtgaat cgataaggat 1381 ccgccccwct ccctccccys scsmtaacgt tactggccga agccgcttgg aataaggccg 1441 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc 1501 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa 1561 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag 1621 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg 1681 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg 1741 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa 1801 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg 1861 gtacacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca 1921 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca 1981 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa 2041 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga 2101 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca 2161 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact 2221 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg 2281 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca 2341 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt 2401 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg 2461 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc 2521 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca 2581 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt 2641 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgca 2701 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 2761 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 2821 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 2881 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 2941 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 3001 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 3061 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 3121 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 3181 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 3241 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct gcctgcaggt 3301 ttgtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc 3361 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca 3421 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 3481 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg 3541 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat 3601 cagatatcca ctgacctttg aatggtgcta caagctagta ccagttgagc aagagaaggt 3661 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat 3721 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca 3781 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc 3841 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 3901 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 3961 acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct 4021 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga 4081 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt 4141 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg 4201 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg 4261 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg 4321 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 4381 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 4441 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 4501 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 4561 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 4621 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt 4681 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg 4741 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 4801 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 4861 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 4921 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 4981 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt 5041 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc 5101 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac 5161 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc 5221 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg 5281 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag 5341 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg 5401 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg 5461 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac 5521 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc 5581 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 5641 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 5701 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 5761 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 5821 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 5881 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 5941 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 6001 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 6061 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 6121 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 6181 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 6241 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 6301 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 6361 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 6421 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 6481 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 6541 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 6601 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 6661 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 6721 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 6781 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 6841 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 6901 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 6961 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 7021 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 7081 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 7141 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 7201 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 7261 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 7321 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 7381 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 7441 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 7501 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 7561 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 7621 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 7681 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 7741 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 7801 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 7861 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 7921 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 7981 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat 8041 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg 8101 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag 8161 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg 8221 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa 8281 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa 8341 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 8401 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 8461 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 8521 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 8581 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 8641 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 8701 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 8761 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 8821 gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc 8881 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa 8941 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag 9001 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac 9061 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg 9121 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg 9181 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt 9241 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc 9301 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac 9361 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata 9421 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt 9481 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg 9541 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat 9601 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg 9661 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca 9721 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct 9781 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg 9841 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac 9901 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact 9961 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg 10021 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt 10081 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg 10141 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata 10201 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat 10261 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga 10321 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt 10381 cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat 10441 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga 10501 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa 10561 ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattaacc aaggcagccc 10621 tggttaac //