LOCUS Exported 11063 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Barcoded lentiviral vector to express TBX5 in mammalian cells under control of EF1a promoter. Barcoded for transcript detection in single cell RNA-seq.. ACCESSION . VERSION . KEYWORDS EF1a_TBX5_P2A_Hygro_Barcode SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11063) AUTHORS Parekh U, Wu Y, Zhao D, Worlikar A, Shah N, Zhang K, Mali P TITLE Mapping Cellular Reprogramming via Pooled Overexpression Screens with Paired Fitness and Single-Cell RNA-Sequencing Readout. JOURNAL Cell Syst. 2018 Nov 12. pii: S2405-4712(18)30433-2. doi: 10.1016/j.cels.2018.10.008. PUBMED 30448000 REFERENCE 2 (bases 1 to 11063) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11063 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1680..1913 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2098..2142 /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 2440..2557 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2693..3871 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2924..3862 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3819..3839 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind 3888..3907 /label=T7 /note="T7 promoter, forward primer" promoter 3888..3906 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 3964..3973 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5536..5592 /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 5593..6618 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" protein_bind 6810..6843 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." LTR 6868..7048 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(7074..7091) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 7080..7304 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 7350..7778 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7437..7456) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7647..7668 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7787..7807) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7792..8121 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7972..8107 /label=SV40 ori /note="SV40 origin of replication" primer_bind 8034..8053 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 8169..8216 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 8235..8609 /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 8739..8860 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8776..8795) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8830..8849 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8909..8925) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8909..8925) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8922..8944) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8933..8949 /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(8957..8987) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9002..9023 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(9140..9157) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(9311..9899) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(9391..9410) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(10070..10930) /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 10693..10712 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10931..11035) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 2581 agatccagtt tggttaatta atcgataagc tttgcaaaga tggataaagt tttaaacaga 2641 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg 2701 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt 2761 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt 2821 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc 2881 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg 2941 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac 3001 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc 3061 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg 3121 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa 3181 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag 3241 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg 3301 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc 3361 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc 3421 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg 3481 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg 3541 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc 3601 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc 3661 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc 3721 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt 3781 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca 3841 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac 3901 tatagggaga cccaagctgg ctaggtaagc ttgatatcga attcctgcag cccgggggat 3961 cccgccacca tggccgacgc agacgagggc tttggcctgg cgcacacgcc tctggagcct 4021 gacgcaaaag acctgccctg cgattcgaaa cccgagagcg cgctcggggc ccccagcaag 4081 tccccgtcgt ccccgcaggc cgccttcacc cagcagggca tggagggaat caaagtgttt 4141 ctccatgaaa gagaactgtg gctaaaattc cacgaagtgg gcacggaaat gatcataacc 4201 aaggctggaa ggcggatgtt tcccagttac aaagtgaagg tgacgggcct taatcccaaa 4261 acgaagtaca ttcttctcat ggacattgta cctgccgacg atcacagata caaattcgca 4321 gataataaat ggtctgtgac gggcaaagct gagcccgcca tgcctggccg cctgtacgtg 4381 cacccagact cccccgccac cggggcgcat tggatgaggc agctcgtctc cttccagaaa 4441 ctcaagctca ccaacaacca cctggaccca tttgggcata ttattctaaa ttccatgcac 4501 aaataccagc ctagattaca catcgtgaaa gcggatgaaa ataatggatt tggctcaaaa 4561 aatacagcgt tctgcactca cgtctttcct gagactgcgt ttatagcagt gacttcctac 4621 cagaaccaca agatcacgca attaaagatt gagaataatc cctttgccaa aggatttcgg 4681 ggcagtgatg acatggagct gcacagaatg tcaagaatgc aaagtaaaga atatcccgtg 4741 gtccccagga gcaccgtgag gcaaaaagtg gcctccaacc acagtccttt cagcagcgag 4801 tctcgagctc tctccacctc atccaatttg gggtcccaat accagtgtga gaatggtgtt 4861 tccggcccct cccaggacct cctgcctcca cccaacccat acccactgcc ccaggagcat 4921 agccaaattt accattgtac caagaggaaa gaggaagaat gttccaccac agaccatccc 4981 tataagaagc cctacatgga gacatcaccc agtgaagaag attccttcta ccgctctagc 5041 tatccacagc agcagggcct gggtgcctcc tacaggacag agtcggcaca gcggcaagct 5101 tgcatgtatg ccagctctgc gccccccagc gagcctgtgc ccagcctaga ggacatcagc 5161 tgcaacacgt ggccaagcat gccttcctac agcagctgca ccgtcaccac cgtgcagccc 5221 atggacaggc taccctacca gcacttctcc gctcacttca cctcggggcc cctggtccct 5281 cggctggctg gcatggccaa ccatggctcc ccacagctgg gagagggaat gttccagcac 5341 cagacctccg tggcccacca gcctgtggtc agacagtgtg ggcctcagac tggcctgcag 5401 tcccctggca cccttcagcc ccctgagttc ctctactctc atggcgtgcc aaggactcta 5461 tcccctcatc agtaccactc tgtgcacgga gttggcatgg tgccagagtg gagcgacaat 5521 agcttgggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag 5581 aatcctggac cgatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc 5641 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct 5701 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt 5761 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa 5821 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag 5881 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg 5941 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc 6001 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat 6061 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag 6121 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac 6181 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac 6241 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg 6301 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt 6361 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag 6421 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc 6481 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc 6541 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt 6601 ccgagggcaa aggaataagt tagaactatt tcctggctgt tacgcgagtt tgctctagga 6661 attttcctac tgacgggctt gttaaacagt gaaccgcccg ccccacgacc cgcagcgccc 6721 gaccgaaagg agcgcacgac cccatgcatc gaacaaaaga aaagggggga ctggaagggc 6781 taattcactc ccaacgaaga caagatatca taacttcgta tagcatacat tatacgaagt 6841 tatcggctag ctggtccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg 6901 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 6961 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 7021 ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac 7081 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct 7141 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct 7201 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg 7261 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag 7321 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc 7381 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc 7441 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 7501 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 7561 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 7621 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 7681 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt 7741 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag 7801 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 7861 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 7921 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 7981 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 8041 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt 8101 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat 8161 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg 8221 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg 8281 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg 8341 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc 8401 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg 8461 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg 8521 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg 8581 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg 8641 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc 8701 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata 8761 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 8821 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga 8881 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 8941 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 9001 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 9061 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 9121 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 9181 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 9241 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 9301 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 9361 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 9421 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 9481 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 9541 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 9601 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 9661 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 9721 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 9781 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 9841 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 9901 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 9961 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 10021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 10081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 10141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 10201 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 10261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 10321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 10381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 10441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 10501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 10561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 10621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 10681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 10741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 10801 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 10861 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 10921 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 10981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 11041 ttccccgaaa agtgccacct gac //