LOCUS       sequence-413610-       15064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPRi for STK3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15064)
  AUTHORS   Ma X, Mandausch FJ, Wu Y, Sahoo VK, Ma W, Leoni G, Hostiuc M, 
            Wintgens JP, Qiu J, Kannaiyan N, Rossner MJ, Wehr MC
  TITLE     Comprehensive split TEV based protein-protein interaction screening 
            reveals TAOK2 as a key modulator of Hippo signalling to limit 
            growth.
  JOURNAL   Cell Signal. 2024 Jan;113:110917. doi: 
            10.1016/j.cellsig.2023.110917. Epub 2023 Oct 7.
  PUBMED    37813295
REFERENCE   2  (bases 1 to 15064)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Signal."; date: "2024-01"; volume: "113:110917. doi"; pages: " 
            10.1016/j.cellsig.2023.110917. Epub 2023 Oct 7"
COMMENT     Sequence Label: pLV-NeoR_dCas9-Sniper-KRAB_sgSTK3i_#2 (NeoR)
FEATURES             Location/Qualifiers
     source          1..15064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             248..268
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             275..328
                     /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."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             323..1123
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(383..402)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     993..1012
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(1139..1172)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1197..1785
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1250..1270)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1668..1679)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1788..1804)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1789..1805)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2310..2490
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(2516..2533)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2522..2746
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2792..3220
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2879..2898)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3089..3110
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3229..3249)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3234..3563
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3414..3549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3476..3495
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3611..3658
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3677..4051
                     /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    4181..4302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4218..4237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4272..4291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4351..4367)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4351..4367)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4364..4386)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4375..4391
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4399..4405,4406..4423,4424..4429))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4444..4465
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4582..4599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4753..5341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4833..4852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5512..6303,6304..6372))
                     /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     6135..6154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6373..6477)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6552..6571)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6743..7122
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7124..7322
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7277..7297
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7340..7520
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7567..7692
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8185..8418
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8603..8647
                     /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    8945..9062
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(9106..9139)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_RNA        complement(9150..9225)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(9246..9494)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(9304..9323)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(9474..9494)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        9527..10738
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     10575..10594
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      10767..10776
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10776..10841
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             10842..10889
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             15015..15035
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(15051..15064,1..181)
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
ORIGIN
        1 cttcaaggat gtgtttgtgg acttcaccag ggaggagtgg aagctgctgg acactgctca
       61 gcagatcctg tacagaaatg tgatgctgga gaactataag aacctggttt ccttgggtta
      121 tcagcttact aagccagatg tgatcctccg gttggagaag ggagaagagc cctggctggt
      181 ggagagagaa attcaccaag agacccatcc tgattcagag actgcatttg aaatcaaatc
      241 atcagttccg aaaaagaaac gcaaagttgc tagcgagggc agaggaagtc ttctaacatg
      301 cggtgacgtg gaggagaatc ccggccctat gattgaacaa gatggattgc acgcaggttc
      361 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
      421 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
      481 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
      541 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
      601 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
      661 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
      721 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
      781 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
      841 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
      901 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
      961 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     1021 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     1081 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaaccggtt gataatagat
     1141 aacttcgtat agcatacatt atacgaagtt atgaattcga tatcaagctt atcgataatc
     1201 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     1261 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     1321 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     1381 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     1441 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     1501 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     1561 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     1621 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     1681 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     1741 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     1801 ctcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
     1861 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
     1921 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     1981 aggggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
     2041 tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca
     2101 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag
     2161 aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg
     2221 atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca
     2281 tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct
     2341 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     2401 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     2461 ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg
     2521 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2581 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2641 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2701 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2761 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2821 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2881 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2941 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3001 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3061 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3121 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3181 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3241 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3301 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3361 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3421 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3481 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3541 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3601 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     3661 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
     3721 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
     3781 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
     3841 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
     3901 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
     3961 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
     4021 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
     4081 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4141 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4201 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4261 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4321 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4381 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4441 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4501 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4561 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4621 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4681 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5161 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5581 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5701 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5941 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6001 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6181 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6361 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6481 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     6541 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     6601 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     6661 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     6721 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     6781 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     6841 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6901 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6961 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     7021 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     7081 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     7141 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     7201 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     7261 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     7321 agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     7381 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7441 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7501 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     7561 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     7621 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     7681 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     7741 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     7801 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     7861 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     7921 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     7981 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     8041 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     8101 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     8161 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     8221 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     8281 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     8341 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     8401 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     8461 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     8521 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     8581 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     8641 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     8701 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     8761 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     8821 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     8881 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     8941 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     9001 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     9061 ttcgggttta ttacagggac agcagagatc cagtttggtt aattaataac ttcgtatagc
     9121 atacattata cgaagttatt aaaaaaaaag caccgactcg gtgccacttt ttcaagttga
     9181 taacggacta gccttatttt aacttgctat ttctagctct aaaaccagag ggaaactctg
     9241 ggaacggtgt ttcgtccttt ccacaagata tataaagcca agaaatcgaa atactttcaa
     9301 gttacggtaa gcatatgata gtccatttta aaacataatt ttaaaactgc aaactaccca
     9361 agaaattatt actttctacg tcacgtattt tgtactaata tctttgtgtt tacagtcaaa
     9421 ttaattccaa ttatctctct aacagccttg tatcgtatat gcaaatatga aggaatcatg
     9481 ggaaataggc cctcttaatt aattaacccg tgtcggctcc agatctggcc tccgcgccgg
     9541 gttttggcgc ctcccgcggg cgcccccctc ctcacggcga gcgctgccac gtcagacgaa
     9601 gggcgcagcg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     9661 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     9721 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     9781 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     9841 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     9901 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     9961 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
    10021 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
    10081 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
    10141 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa
    10201 agctcttatt cgggtgagat gggctggggc accatctggg gaccctgacg tgaagtttgt
    10261 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt
    10321 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt
    10381 tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct
    10441 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga
    10501 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta
    10561 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt
    10621 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca
    10681 gtgttagact agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacga
    10741 agcttgggct gcaggtcgac tctagagcca ccatggacta caaagaccat gacggtgatt
    10801 ataaagatca tgacatcgac tacaaggatg acgatgacaa gaagcgacct gccgccacaa
    10861 agaaggctgg acaggctaag aagaagaaag aagcgtccga caagaagtac agcatcggcc
    10921 tggccatcgg caccaacagc gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
    10981 gcaagaagtt caaggtgctg ggcaacaccg accgccacag catcaagaag aacctgatcg
    11041 gcgctctgct gttcgacagc ggagagaccg ccgaggccac acgcctgaag cgcaccgcca
    11101 gacgccgcta caccagacgc aagaaccgca tctgctacct gcaggagatc ttcagcaacg
    11161 agatggccaa ggtggacgac agcttcttcc accgcctgga ggagagcttc ctggtggagg
    11221 aggacaagaa gcacgagcgc caccccatct tcggcaacat cgtggacgag gtggcctacc
    11281 acgagaagta ccccaccatc taccacctgc gcaagaagct ggtggacagc accgacaagg
    11341 ccgacctgcg cctgatctac ctggccctgg cccacatgat caagttccgc ggccacttcc
    11401 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
    11461 tgcagaccta caaccagctg ttcgaggaga accccatcaa cgccagcggc gtggacgcca
    11521 aggccatcct gagcgcccgc ctgagcaaga gccgccgcct ggagaacctg atcgcccagc
    11581 tgcccggcga gaagaagaac ggcctgttcg gcaacctgat cgccctgagc ctgggcctga
    11641 cccccaactt caagagcaac ttcgacctgg ccgaggacgc caagctgcag ctgagcaagg
    11701 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
    11761 tgttcctggc cgccaagaac ctgagcgacg ccatcctgct gagcgacatc ctgcgcgtga
    11821 acaccgagat caccaaggcc cccctgagcg ccagcatgat caagcgctac gacgagcacc
    11881 accaggacct gaccctgctg aaggccctgg tgcgccagca gctgcccgag aagtacaagg
    11941 agatcttctt cgaccagagc aagaacggct acgccggcta catcgacggc ggcgccagcc
    12001 aggaggagtt ctacaagttc atcaagccca tcctggagaa gatggacggc accgaggagc
    12061 tgctggtgaa gctgaaccgc gaggacctgc tgcgcaagca gcgcaccttc gacaacggca
    12121 gcatccccca ccagatccac ctgggcgagc tgcacgccat cctgcgccgc caggaggact
    12181 tctacccctt cctgaaggac aaccgcgaga agatcgagaa gatcctgacc ttccgcatcc
    12241 cctactacgt gggccccctg gcccgcggca acagccgctt cgcctggatg acccgcaaga
    12301 gcgaggagac catcaccccc tggaacttcg aggaggtggt ggacaagggc gccagcgccc
    12361 agagcttcat cgagcgcatg accaacttcg acaagaacct gcccaacgag aaggtgctgc
    12421 ccaagcacag cctgctgtac gagtacttca ccgtgtacaa cgagctgacc aaggtgaagt
    12481 acgtgaccga gggcatgcgc aagcccgcct ccctgagcgg cgagcagaag aaggccatcg
    12541 tggacctgct gttcaagacc aaccgcaagg tgaccgtgaa gcagctgaag gaggactact
    12601 tcaagaagat cgagtgcttc gacagcgtgg agatcagcgg cgtggaggac cgcttcaacg
    12661 ccagcctggg cacctaccac gacctgctga agatcatcaa ggacaaggac ttcctggaca
    12721 acgaggagaa cgaggacatc ctggaggaca tcgtgctgac cctgaccctg ttcgaggacc
    12781 gcgagatgat cgaggagcgc ctgaagacct acgcccacct gttcgacgac aaggtgatga
    12841 agcagctgaa gcgccgccgc tacaccggct ggggccgcct gagccgcaag cttatcaacg
    12901 gcatccgcga caagcagagc ggcaagacca tcctggactt cctgaagagc gacggcttcg
    12961 ccaaccgcaa cttcatgcag ctgatccacg acgacagcct gaccttcaag gaggacatcc
    13021 agaaggccca ggtgagcggc cagggcgaca gcctgcacga gcacatcgcc aacctggccg
    13081 gcagccccgc catcaagaag ggcatcctgc agaccgtgaa ggtggtggac gagctggtga
    13141 aggtgatggg ccgccacaag cccgagaaca tcgtgatcga gatagcccgc gagaaccaga
    13201 ccacccagaa gggccagaag aacagccgcg agcgcatgaa gcgcatcgag gagggcatca
    13261 aggagctggg cagccagatc ctgaaggagc accccgtgga gaacacccag ctgcagaacg
    13321 agaagctgta cctgtactac ctgcagaacg gccgcgacat gtacgtggac caggagctgg
    13381 acatcaaccg cctgagcgac tacgacgtgg acgccatcgt gccccagagc ttcctgaagg
    13441 acgacagcat cgacaacaag gtgctgaccc gcagcgacaa gaaccgcggc aagagcgaca
    13501 acgtgcccag cgaggaggtg gtgaagaaga tgaagaacta ctggcgccag ctgctgaacg
    13561 ccaatctgat cacccagcgc aagttcgaca acctgaccaa ggccgagcgc ggcggcctga
    13621 gcgagctgga caaggccggc ttcatcaagc gccagctggt ggagacccgc cagatcacca
    13681 agcacgtggc ccagatcctg gacagccgca tgaacaccaa gtacgacgag aacgacaagc
    13741 tgatccgcga ggtgaaggtg atcaccctga agagcaagct ggtgagcgac ttccgcaagg
    13801 acttccagtt ctacaaggtg cgcgagatca acaactacca ccacgcccac gacgcctacc
    13861 tgaacgccgt ggtgggcacc gccctgatca agaagtaccc caagctggag agcgagttcg
    13921 tgtacggcga ctacaaggta tacgacgtgc gcaagatgat cgctaagagc gagcaggaga
    13981 tcggcaaggc caccgccaag tacttcttct acagcaacat catgaacttc ttcaagaccg
    14041 agatcaccct ggccaacggc gagatccgca agcgccccct gatcgagacc aacggcgaga
    14101 ccggcgagat cgtgtgggac aagggccgcg acttcgccac cgtgcgcaag gtgctgagca
    14161 tgccccaggt gaacatcgtg aagaagaccg aggtgcagac cggcggcttc agcaaggaga
    14221 gcatcctgcc caagcgcaac agcgacaagc tgatcgcccg caagaaggac tgggacccca
    14281 agaagtacgg cggcttcgac agccccaccg tggcctacag cgtgctggtg gtggccaagg
    14341 tggagaaggg caagagcaag aagctgaaga gcgtgaagga gctgctgggc atcaccatca
    14401 tggagcgcag cagcttcgag aagaacccca tcgacttcct ggaggccaag ggctacaagg
    14461 aggtgaagaa ggacctgatc atcaagctgc ccaagtacag cctgttcgag ctggagaacg
    14521 gccgcaagcg catgctggcc agcgccggcg agctgcagaa gggcaacgag ctggccctgc
    14581 ccagcaagta cgtgaacttc ctgtacctgg ccagccacta cgagaagctg aagggcagcc
    14641 ccgaggacaa cgagcagaag cagctgttcg tggagcagca caagcactac ctggacgaga
    14701 tcatcgagca gatcagcgag ttcagcaagc gcgtgatcct ggccgacgcc aacctggaca
    14761 aggtgctgag cgcctacaac aagcaccgcg acaagcccat ccgcgagcag gccgagaaca
    14821 tcatccacct gttcaccctg accaacctgg gcgcccccgc cgccttcaag tacttcgaca
    14881 ccaccatcga ccgcaagcgc tacaccagca ccaaggaggt gctggacgcc accctgatcc
    14941 accagagcat caccggtctg tacgagaccc gcatcgacct gagccagctg ggcggcgacg
    15001 gcggctccgg acctccaaag aaaaagagaa aagtaggagg tggaggtagc cggacactgg
    15061 tgac
//
