LOCUS pHACK-GAL4_ZpQFD 9879 bp DNA circular SYN 26-APR-2025 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 9879) AUTHORS Junjie Luo TITLE Direct Submission JOURNAL Exported Apr 26, 2025 from SnapGene 8.0.3 https://www.snapgene.com FEATURES Location/Qualifiers source 1..9879 /mol_type="other DNA" /organism="recombinant plasmid" protein_bind 41..74 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" /note="color: #31849b; direction: RIGHT" protein_bind complement(88..157) /label=attB /bound_moiety="phage phi-C31 integrase" /note="attB site for the phi-C31 integrase (Groth et al., 2000)" /note="color: #31849b; direction: LEFT" protein_bind 88..157 /label=attB /bound_moiety="phage phi-C31 integrase" /note="attB site for the phi-C31 integrase (Groth et al., 2000)" /note="color: #31849b" primer_bind complement(88..110) /label=AD2_attB_typing_f2 /note="color: black; sequence: GCGCGTACTCCACCTCACCCATC; added: 2020-02-17" primer_bind complement(137..157) /label=AD2_attB_typing_f /note="color: black; sequence: CTCGAAGCCGCGGTGCGGGTG; added: 2020-02-17" misc_feature join(175..205,206..313) /label=P element 5' end (Potter) /note="first 139 bp of P element 5' end (586 bp)" /note="This feature has 2 segments: 1: 175 .. 205 / #ffcc99 / inverted repreat 2: 206 .. 313 / #ffcc99" protein_bind 326..359 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" /note="color: #31849b; direction: RIGHT" misc_feature 399..1555 /label=GAL4_HA1 /note="color: #a6acb3; direction: RIGHT" CDS 1568..1621 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /note="color: #cc99b2 Cleavage site after base 1618" /translation="EGRGSLLTCGDVEENPGP" CDS join(1628..1807,1808..2356) /codon_start=1 /label=Zp-::QFDBD /note="This feature has 2 segments: 1: 1628 .. 1807 / #993366 / Zp- 2: 1808 .. 2356 / #993366 / QFDBD" /translation="MLEIRAAFLRQRNTALRTEVAELEQEVQRLENEVSQYETRYGPLG GGKASGGGGGGGGDVMPPKRKTLNAAAEANAHADGHADGNADGHVANTAASSNNARFAD LTNIDTPGLGPTTTTLLVEPARSKRQRVSRACDQCRAAREKCDGIQPACFPCVSQGRSC TYQASPKKRGVQTGYIRTLELALAWMFENVARSEDALHNLLVRDAGQGSALLVGKDSPA AERLHARWATSRVNKSITRLL" CDS 1628..1802 /codon_start=1 /label=ZpGAL4DBD /label=ORF frame 1 /note="color: #993366" /translation="MLEIRAAFLRQRNTALRTEVAELEQEVQRLENEVSQYETRYGPLG GGKASGGGGGGGG" primer_bind 1628..1642 /label=Zp_primer_f /note="color: black; sequence: ATCGACAGCCGAATTCAACATGCTGGAGATCCGC; added: 2020-12-27" CDS 1808..2356 /codon_start=1 /label=QF2 /note="color: #993366" /translation="MPPKRKTLNAAAEANAHADGHADGNADGHVANTAASSNNARFADL TNIDTPGLGPTTTTLLVEPARSKRQRVSRACDQCRAAREKCDGIQPACFPCVSQGRSCT YQASPKKRGVQTGYIRTLELALAWMFENVARSEDALHNLLVRDAGQGSALLVGKDSPAA ERLHARWATSRVNKSITRLL" polyA_signal 2375..2620 /label=hsp70 polyA /note="color: #a6acb3; direction: RIGHT" misc_feature join(2625,2626..2675,2676..2681,2682..3841,3842..3858) /label=[AD2_B^{Bxb1}] /note="This forward directional feature has 5 segments: 1: 2625 .. 2625 / #ffffff 2: 2626 .. 2675 / #31849b / attB[Bxb1] 3: 2676 .. 2681 / #ffffff 4: 2682 .. 3841 / #00ffff / 2xr4-CFP 5: 3842 .. 3858 / #ffffff" misc_feature complement(2626..2675) /label=attB(Bxb1) /note="http://flybase.org/reports/FBto0000359 The attB(Bxb1) site (5'-TCGGCCGGCTTGTCGACGACGgcggtctcCGTCGTCAGGATCATCCGGGC-3') is one of the two target sites of the Bxb1:int integrase. Bxb1:int mediates recombination between two non-identical target sites, attP(Bxb1) and attB(Bxb1), each of which contains a short integration core flanked by imperfect inverted repeats. Exchange between the attP(Bxb1)/attB(Bxb1) pair results in the formation of hybrid sites that are no longer a substrate of the Bxb1:int integrase, and thus the exchange reaction driven by Bxb1:int is undirectional (reviewed in PMID:21195181, FBrf0231034). The recombination event catalyzed by Bxb1:int results in genetic modification, the nature of which is influenced by the relative orientation (direct or inverted), and location of the two target sites. The types of possible modification include deletion of DNA, generation of chromosomal rearrangements, integration of DNA into the genome, and replacement of genetic material with that from a donor plasmid using recombination-mediated cassette exchange (RMCE) (reviewed in FBrf0231034)." /note="color: #31849b; direction: LEFT" polyA_signal 2682..2803 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" primer_bind complement(2698..2727) /label=SV40A_seq_f /note="color: black; sequence: CACTGCATTCTAGTTGTGGTTTGTCCAAAC; added: 2021-10-19" CDS complement(join(2810..3523,3524..3526,3527..3529)) /codon_start=1 /product="cyan variant of GFP" /label=CFP /note="mammalian codon-optimized" /note="This feature has 3 segments: 1: 2810 .. 3523 / #00ffff 2: 3524 .. 3526 / #00ffff / 1a 3: 3527 .. 3529 / #00ffff" /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2813..2842) /label=CFP_230_seq_fL /note="color: black; sequence: ATCACTCTCGGCATGGACGAGCTGTACAAG; added: 2021-12-14" primer_bind complement(2813..2835) /label=CFP_230_seq_f /note="color: black; sequence: TCGGCATGGACGAGCTGTACAAG; added: 2021-09-18" primer_bind complement(2827..2855) /label=CFP_224_seq_f /note="color: black; sequence: GACCGCCGCCGGGATCACTCTCGGCATGG; added: 2021-10-19" primer_bind complement(2855..2884) /label=CFP_215_seq_f /note="color: black; sequence: CGCGATCACATGGTCCTGCTGGAGTTCGTG; added: 2021-10-19" primer_bind 3392..3414 /label=CFP_45_seq_r /note="color: black; sequence: CTTCAGGGTCAGCTTGCCGTAGG; added: 2021-09-18" promoter complement(join(3540..3625,3653..3658,3659..3664)) /label=hsp70 mini /note="Drosophila melanogaster hsp70Bb promoter" /note="This reverse directional feature has 3 segments: 1: 3540 .. 3625 / #ffffff / align to 3xP3 2: 3653 .. 3658 / #cc99ff / TATA box 3: 3659 .. 3664 / #ffffff" promoter complement(join(3684..3699,3704..3719,3724..3739, 3744..3759)) /label=r4 promoter /note="This reverse directional feature has 4 segments: 1: 3684 .. 3699 / #ffffff 2: 3704 .. 3719 / #ffffff 3: 3724 .. 3739 / #ffffff 4: 3744 .. 3759 / #ffffff" promoter complement(join(3766..3781,3786..3801,3806..3821, 3826..3841)) /label=r4 promoter /note="This reverse directional feature has 4 segments: 1: 3766 .. 3781 / #ffffff 2: 3786 .. 3801 / #ffffff 3: 3806 .. 3821 / #ffffff 4: 3826 .. 3841 / #ffffff" protein_bind 3864..3897 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." /note="color: #31849b; direction: RIGHT" misc_feature 3908..5276 /label=GAL4_HA2 /note="color: #a6acb3; direction: RIGHT" CDS 4932..5276 /codon_start=1 /gene="Saccharomyces cerevisiae GAL4 (truncated)" /product="activation domain of the GAL4 transcriptional activator" /label=GAL4 activation domain /note="color: #993366" /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD EDTPPNPKKE" protein_bind 5286..5319 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" /note="color: #31849b; direction: RIGHT" promoter 5335..6131 /label=dU6-1 promoter /note="RNA polymerase III promoter for Drosophila U6-1 snRNA (Port et al., 2014)" /note="color: #ffffff; direction: RIGHT" misc_RNA 6151..6226 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" /note="color: #00ccff; direction: RIGHT" promoter 6227..6647 /label=dU6-3 promoter /note="RNA polymerase III promoter for Drosophila U6-3 snRNA (Port et al., 2014)" /note="color: #ffffff; direction: RIGHT" promoter 6233..6647 /label=U6:3 promoter /note="color: #ffffff; direction: RIGHT" misc_RNA 6668..6743 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" /note="color: #00ccff; direction: RIGHT" misc_feature join(7003..7134,7135..7165) /label=P element 3' end (Potter) /note="last 163 bp of P element 3' end (233 bp) Editing Transgenic DNA Components by Inducible Gene Replacement in Drosophila melanogaster Editing Transgenic DNA Components by InducibleGene Replacement in Drosophila melanogaster Chun-Chieh Lin and Christopher J. Potter1 The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205 Editing Transgenic DNA Components by Inducible Gene Replacement in Drosophila melanogaster To generate a versatile construct with multiple synthetic com-ponents, we de novo designed the donor vector for DNA syn-thesis (DNA2.0, Newark, CA). The construct contains 59 and 39P-element sequences, attB sequence, T2A, origin of replication,and ampicillin resistance sequence. " /note="This feature has 2 segments: 1: 7003 .. 7134 / #ffcc99 2: 7135 .. 7165 / #ffcc99 / inverted repeat" protein_bind complement(7176..7209) /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" /note="color: #31849b; direction: LEFT" primer_bind complement(7679..7696) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" /note="color: black; sequence: TGTAAAACGACGGCCAGT" primer_bind complement(7679..7695) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: LEFT" rep_origin complement(7965..8553) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" primer_bind complement(8045..8064) /label=pBR322ori-F /note="pBR322 origin, forward primer" /note="color: black; sequence: GGGAAACGCCTGGTATCTTT" CDS complement(join(8675..9466,9467..9535)) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 8675 .. 9466 / #ccffcc 2: 9467 .. 9535 / #ccffcc / signal sequence Cleavage site after base 9466" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 9298..9317 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" /note="color: black; sequence: ATAATACCGCGCCACATAGC" promoter complement(9536..9627) /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: LEFT" primer_bind 9669..9688 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" /note="color: black; sequence: ATCGCGAGCCCATTTATACC" primer_bind 9694..9713 /label=pENTR-R /note="pENTR vectors, reverse primer" /note="color: black; sequence: ATGGCTCATAACACCCCTTG" ORIGIN 1 gggctaatta tggggtgtcg cccttattcg actctatagt gaagttccta ttctctagaa 61 agtataggaa cttctgaagt ggggcgggat gggtgaggtg gagtacgcgc ccggggagcc 121 caagggcacg ccctggcacc cgcaccgcgg cttcgagtac aagccgaagg cgcgcatgat 181 gaaataacat aaggtggtcc cgtcgaaagc cgaagcttac cgaagtatac acttaaattc 241 agtgcacgtt tgcttgttga gaggaaaggt tgtgtgcgga cgaatttttt tttgaaaaca 301 ttaaccctta cgtaacgcga gatctgaagt tcctattctc tagaaagtat aggaacttct 361 agggataaca gggtaataga tctggatcct aatgatgaca tgcgatattt gccgacttaa 421 aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga 481 gtgtcgctac tctcccaaaa ccaaaaggtc tccgctgact agggcacatc tgacagaagt 541 ggaatcaagg ctagaaagac tggaacagct atttctactg atttttcctc gagaagacct 601 tgacatgatt ttgaaaatgg attctttaca ggatataaaa gcattgttaa caggattatt 661 tgtacaagat aatgtgaata aagatgccgt cacagataga ttggcttcag tggagactga 721 tatgcctcta acattgagac agcatagaat aagtgcgaca tcatcatcgg aagagagtag 781 taacaaaggt caaagacagt tgactgtatc gattgactcg gcagctcatc atgataactc 841 cacaattccg ttggatttta tgcccaggga tgctcttcat ggatttgatt ggtctgaaga 901 ggatgacatg tcggatggct tgcccttcct gaaaacggac cccaacaata atgggttctt 961 tggcgacggt tctctcttat gtattcttcg atctattggc tttaaaccgg aaaattacac 1021 gaactctaac gttaacaggc tcccgaccat gattacggat agatacacgt tggcttctag 1081 atccacaaca tcccgtttac ttcaaagtta tctcaataat tttcacccct actgccctat 1141 cgtgcactca ccgacgctaa tgatgttgta taataaccag attgaaatcg cgtcgaagga 1201 tcaatggcaa atccttttta actgcatatt agccattgga gcctggtgta tagaggggga 1261 atctactgat atagatgttt tttactatca aaatgctaaa tctcatttga cgagcaaggt 1321 cttcgagtca ggttccataa ttttggtgac agccctacat cttctgtcgc gatatacaca 1381 gtggaggcag aaaacaaata ctagctataa ttttcacagc ttttccataa gaatggccat 1441 atcattgggc ttgaataggg acctcccctc gtccttcagt gatagcagca ttctggaaca 1501 aagacgccga atttggtggt ctgtctactc ttgggagatc caattgtccc tgcttggatc 1561 ctcgcgtgag ggccgcggca gcctgctgac ctgcggcgat gtggaggaga accccgggcc 1621 ccctaggatg ctggagatcc gcgccgcctt cctgcgccag cgcaacaccg ccctgcgcac 1681 cgaggtggcc gagctggagc aggaggtgca gcgcctggag aacgaggtga gccagtacga 1741 gacccgctac ggccccctgg gcggcggcaa ggctagcgga ggaggtggtg gaggtggagg 1801 tgacgtgatg ccacccaagc gcaaaacgct taacgctgcg gctgaggcta acgctcatgc 1861 cgacggacac gccgacggaa acgccgacgg acacgtggcc aatacggccg cgtcctcgaa 1921 taatgcgagg ttcgctgatc tcactaacat cgatactccg ggtctgggac ccacaactac 1981 gaccctgctc gtggaaccag cacgctcaaa gcgtcaacga gtgtcccgcg catgcgacca 2041 gtgccgtgca gcccgagaga aatgcgacgg aatacagcct gcgtgtttcc cgtgcgtttc 2101 ccagggaagg tcctgcactt atcaggcttc gccgaaaaag aggggagttc aaaccggtta 2161 tattcgtacg ctggagctcg ccctcgcctg gatgtttgaa aatgtcgcgc gttccgaaga 2221 tgccttgcat aacctcctcg tccgtgacgc cggacaagga tcagctctgc tcgttggtaa 2281 agattcgccg gctgccgagc gactccatgc ccgttgggct actagccgtg tcaataagag 2341 cattacccgc ctcctctaaa agcttatcga taccgtcgac taaagccaaa tagaaaatta 2401 ttcagttcct ggcttaagtt tttaaaagtg atattattta tttggttgta accaaccaaa 2461 agaatgtaaa taactaatac ataattatgt tagttttaag ttagcaacaa attgatttta 2521 gctatattag ctacttggtt aataaataga atatatttat ttaaagataa ttgcgttttt 2581 attgtcaggg agtgagtttg cttaaaaact cgtttagatc catatgcccg gatgatcctg 2641 acgacggaga ccgccgtcgt cgacaagccg gccgagcatg ctaagataca ttgatgagtt 2701 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc 2761 tattgcttta tttgtaacca ttataagctg caataaacaa gttaccggtt tacttgtaca 2821 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat 2881 cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg 2941 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca 3001 cgctgccgtc ctcgatgttg tggcggatct tgaagtgggc cttgatgccg ttcttctgct 3061 tgtcggcggt gatatagacg ttgtggctga tgtagttgta ctccagcttg tgccccagga 3121 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc 3181 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtac 3241 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt 3301 cggggtagcg gctgaagcac tgcacgcccc aggtcagggt ggtcacgagg gtgggccagg 3361 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat 3421 cgccctcgcc ctcgccggac acgctgaacc tgtggccgtt tacgtcgccg tccagctcga 3481 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg ttgacgcgtc 3541 cgattgttta gcttgttcag ctgcgcttgt ttatttgctt agctttcgct tagcgacgtg 3601 ttcactttgc ttgtttgaat tgaattgtcg ctccgtagac gaagcgcctc tatttatact 3661 ccggcgtacg tagagtctcc gctcgcctga ttattttaat ctccgcctga ttattttaat 3721 gaccgcctga ttattttaat ctacgcctga ttattttaaa tgcatcgcct gattatttta 3781 atctccgcct gattatttta atgaccgcct gattatttta atctacgcct gattatttta 3841 acgggtcacg gaagttagac gtcataactt cgtatagcat acattatacg aagttataga 3901 tcactagtca tggcatcatt gaaacagcaa ggctcttaca agttttcaca aaaatctatg 3961 aactagacaa aacagtaact gcagaaaaaa gtcctatatg tgcaaaaaaa tgcttgatga 4021 tttgtaatga gattgaggag gtttcgagac aggcaccaaa gtttttacaa atggatattt 4081 ccaccaccgc tctaaccaat ttgttgaagg aacacccttg gctatccttt acaagattcg 4141 aactgaagtg gaaacagttg tctcttatca tttatgtatt aagagatttt ttcactaatt 4201 ttacccagaa aaagtcacaa ctagaacagg atcaaaatga tcatcaaagt tatgaagtta 4261 aacgatgctc catcatgtta agcgatgcag cacaaagaac tgttatgtct gtaagtagct 4321 atatggacaa tcataatgtc accccatatt ttgcctggaa ttgttcttat tacttgttca 4381 atgcagtcct agtacccata aagactctac tctcaaactc aaaatcgaat gctgagaata 4441 acgagaccgc acaattatta caacaaatta acactgttct gatgctatta aaaaaactgg 4501 ccacttttaa aatccagact tgtgaaaaat acattcaagt actggaagag gtatgtgcgc 4561 cgtttctgtt atcacagtgt gcaatcccat taccgcatat cagttataac aatagtaatg 4621 gtagcgccat taaaaatatt gtcggttctg caactatcgc ccaataccct actcttccgg 4681 aggaaaatgt caacaatatc agtgttaaat atgtttctcc tggctcagta gggccttcac 4741 ctgtgccatt gaaatcagga gcaagtttca gtgatctagt caagctgtta tctaaccgtc 4801 caccctctcg taactctcca gtgacaatac caagaagcac accttcgcat cgctcagtca 4861 cgccttttct agggcaacag caacagctgc aatcattagt gccactgacc ccgtctgctt 4921 tgtttggtgg cgccaatttt aatcaaagtg ggaatattgc tgatagctca ttgtccttca 4981 ctttcactaa cagtagcaac ggtccgaacc tcataacaac tcaaacaaat tctcaagcgc 5041 tttcacaacc aattgcctcc tctaacgttc atgataactt catgaataat gaaatcacgg 5101 ctagtaaaat tgatgatggt aataattcaa aaccactgtc acctggttgg acggaccaaa 5161 ctgcgtataa cgcgtttgga atcactacag ggatgtttaa taccactaca atggatgatg 5221 tatataacta tctattcgat gatgaagata ccccaccaaa cccaaaaaaa gagtaataga 5281 ctagtgaagt tcctattctc tagaaagtat aggaacttct agtgatctga attcattttc 5341 aacgtcctcg atagtatagt ggttagtatc cccgcctgtc acgcgggaga ccggggttca 5401 attccccgtc ggggagaatc tgtgattctt tttttttttc ttttactttg ttatataaac 5461 aatttttgtt ttaattgaat ctaatttgcc attgctttta ggaatctcag gcatccagca 5521 agcgtttgtc cgccgaatcg cccatcagtg aagaagatcc tgtggcggct acgaaaatct 5581 ccccggccat gtcggcctcc acctccagcg aaaaacccat cagcgagctg gccacctctg 5641 tgctgaccca ccgctttcca gactccacct cctcacccgg cgaacatggc cttggacgaa 5701 tgcagttgtc gatccgctac agcgcccagc gtcaaaaact agacgtgacc atacacaaaa 5761 tccagaagat accacttcgc gatcccagca atatccccga tccgtatgtt aagctgtatc 5821 tgttgcctgg acgcaccaag gagtcgaaac gcaagacgag cgtgatcaag gacaactgca 5881 accccgtcta cgatgcatcc tttgagtacc tgatttccat tgccgaactc aggcagacgg 5941 aactggaggt gacggtgtgc acccaaaagg gattcctatc cggcggtagt cccatcattg 6001 gcatggtagg tacccgaaag caacccctta gttacagaca cagcgcgtac gtccttcgca 6061 tccttatgat tcccaagtac atattctgca agagtacagt atatatagga aagatatccg 6121 ggtgaacttc gatgtgcagc gtaccacaac gttttagagc tagaaatagc aagttaaaat 6181 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgctcacct 6241 gtgattgctc ctactcaaat acaaaaacat caaattttct gtcaataaag catatttatt 6301 tatatttatt ttacaggaaa gaattccttt taaagtgtat tttaacctat aatgaaaaac 6361 gattaaaaaa aatacataaa ataattcgaa aatttttgaa tagcccaggt tgataaaaat 6421 tcatttcata cgttttataa cttatgcccc taagtatttt ttgaccatag tgtttcaatt 6481 ctacattaat tttacagagt agaatgaaac gccacctact cagccaagag gcgaaaaggt 6541 tagctcgcca agcagagagg gcgccagtgc tcactacttt ttataattct caacttcttt 6601 ttccagactc agttcgtata tatagaccta ttttcaattt aacgtcgtgt attctgagaa 6661 agctggagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg 6721 aaaaagtggc accgagtcgg tgcttttttg cctacctgga gcctgagagt tgttcaataa 6781 aataaaaatg tttcgttttt ttgctttcgc cagtatttat tatttttcat caatatgtat 6841 tcaatttggt atgtatttag taattgtaat atatagacaa tggttttccg ttgacgtaca 6901 tacatctgac gtgtgtttat ttagacataa tagttatgtt ttcacatctt tttaatgttc 6961 gcttaatgcg tatgcttaat taattctgac gtaagctagt tacaaacccc acggacatgc 7021 taagggttaa tcaacaatca tatcgctgtc tcactcagac tcaatacgac actcagaata 7081 ctattccttt cactcgcact tattgcaagc atacgttaag tggatgtctc ttgccgacgg 7141 gaccacctta tgttatttca tcatgaaaat gaagtgaagt tcctatactt tctagagaat 7201 aggaacttct atagtgagtc gaataagggc gacacaaaat ttattctaaa tgcataataa 7261 atactgataa catcttatag tttgtattat attttgtatt atcgttgaca tgtataattt 7321 tgatatcaaa aactgatttt ccctttatta ttttcgagat ttattttctt aattctcttt 7381 aacaaactag aaatattgta tatacaaaaa atcataaata atagatgaat agtttaatta 7441 taggtgttca tcaatcgaaa aagcaacgta tcttatttaa agtgcgttgc ttttttctca 7501 tttataaggt taaataattc tcatatatca agcaaagtga caggcgccct taaatattct 7561 gacaaatgct ctttccctaa actcccccca taaaaaaacc cgccgaagcg ggtttttacg 7621 ttatttgcgg attaacgatt actcgttatc agaaccgccc agggggcccg agcttaagac 7681 tggccgtcgt tttacaacac agaaagagtt tgtagaaacg caaaaaggcc atccgtcagg 7741 ggccttctgc ttagtttgat gcctggcagt tccctactct cgccttccgc ttcctcgctc 7801 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 7861 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 7921 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 7981 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 8041 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 8101 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 8161 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 8221 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 8281 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 8341 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt gggctaacta cggctacact 8401 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 8461 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 8521 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 8581 tctgacgctc agtggaacga cgcgcgcgta actcacgtta agggattttg gtcatgagct 8641 tgcgccgtcc cgtcaagtca gcgtaatgct ctgcttacca atgcttaatc agtgaggcac 8701 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 8761 taactacgat acgggagggc ttaccatctg gccccagcgc tgcgatgata ccgcgagaac 8821 cacgctcacc ggctccggat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 8881 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 8941 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccatcgct acaggcatcg 9001 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 9061 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 9121 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 9181 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 9241 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 9301 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 9361 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 9421 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 9481 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatattct 9541 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 9601 ttgaatgtat ttagaaaaat aaacaaatag gggtcagtgt tacaaccaat taaccaattc 9661 tgaacattat cgcgagccca tttatacctg aatatggctc ataacacccc ttgtttgcct 9721 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt 9781 agcgccgatg gtagtgtggg gactccccat gcgagagtag ggaactgcca ggcatcaaat 9841 aaaacgaaag gctcagtcga aagactgggc ctttcgccc //